Document b50EMqyKrGJN3B5KJ24gxxny

3 3M Environm ental Laboratory E05-0210 Interim R eport#13 Interim Report #13: Analysis of Tennessee River Water Samples Study Title A P (PFBS), Pn a l y s i s o f e r f l u o r o b u t a n e s u l f o n a t e erfluo ro hexanesulfonate (PFHS), a n d Pe r f l u o r o o c t a n e s u l f o n a t e (PFOS) in Wa t e r , So i l , Se d i m e n t , Fi s h , C , V , Sl a m s e g e t a t i o n m a l l Ma m m a l Liv e r a n d Sm a l l M Sa m m a l e r u m Us i n g LC/MS/MS 3M D M Pf o r t h e e c a t u r o n it o r in g r o g r a m Data Requirement EPA TSCA Good Laboratory Practice Standards 40 CFR 792 Author Michelle D. Malinsky, Ph.D 3M Environmental Laboratory Interim Report Completion Date Date of signing Performing Laboratory 3M Environmental Laboratory Building 2-3E-09 935 Bush Ave. St. Paul, MN 55106 Project Identification E05-0210 160 Total Number of Pages E05-0210 Interim R eport#13 Tennessee R iver Page 1 of 160 3M Environm ental Labora tory E05-0210 Interim R eport#13 This page has been reserved for specific country requirements. E05-0210 Interim R eport#13 Tennessee R iver Page 2 of 160 3M Environmental Laboratory E0S-0210 Interim Report#13 GLP Compliance Statement Study Title: Analysis of Perfluorobutanesulfonate (PFBS), Perfluorohexanesulfonate (PFHS), and Perfluorooctanesulfonate (PFOS) in Water, Soil, Sediment, Fish, Clams,Vegetation, Small Mammal Liver and Small Mammal Serum Using LC/MS/MS for the 3M Decatur Monitoring Program Interim Study: Analysis of Tennessee River Water Samples Interim Study Identification Number: E05-210 This study was conducted in compliance with Toxic Substances Control Act (TSCA) Good Laboratory Practice (GLP) Standards, 40 CFR 792, with the exceptions listed below: Michael A. Santoro, ^ Company, Sponsor Representative ' Date E05-0210 Interim Report013 . Tennessee River Page 3 of 160 3M Environmental Laboratory E05-O210 Interim R e p o rta i3 Quality Assurance Statement Study Title: Analysis of Perfluorobutanesulfonate (PFBS), Perfluorohexanesulfonate (PFHS), and Perfluorooctanesulfonate (PFOS) n Water, Soil, Sediment, Fish, Clams,Vegetation, Small Mammal Liver and Small Mammal Serum Using LC/MS/MS for the 3M Decatur Monitoring Program Interim Study: Analysis of Tennessee River Water Samples Interim Study Identification Number: E05-0210, Interim Report #13 This study was audited by the 3M Environmental Laboratory Quality Assurance Unit (QAU), as indicated in the following table. The findings were reported to the study director and laboratory management. Inspection Dates July 27,2005 October 5-7,12,13,2005 Phase In-phase Data/Final Report D ate Reported to Managem ent S tudy D irecto r 11-07-2005 11-07-2005 11-17-2005 11-17-2005 QAU Representative //-/7-or Date E05-0210 Interim Reportai3 Tennessee River Page 4 o f 160 3M Environm ental Laboratory E05-0210 Interim R eport#13 Table of Contents GLP Compliance Statement.............................................................................................................. 3 Quality Assurance Statement............................................................................................................ 4 Table of Contents.............................................................................................................................. 5 List of Tables......................................................................................................................................6 Study Information.............................................................................................................................. 8 Summary and Introduction................................................................................................................. 9 Test Samples................................................................................................................................... 15 Reference Substances..................................................................................................................... 15 Control Substance............................................................................................................................15 Method Summaries..........................................................................................................................16 Sample Collection................................................................................................................ 16 Preparatory and Analytical Methods..................................................................................... 17 Extraction............................................................................................................................. 17 Analysis................................................................................................................................ 17 Analytical Results............................................................................................................................ 18 Calibration............................................................................................................................ 18 Limit of Quantitation (LOQ)................................................................................................... 18 Blanks.................................................................................................................................. 19 Solvent Blank........................................................................................................................19 Method Blank........................................................................................................................19 Trip Blank............................................................................................................................. 21 System Suitability................................................................................................................. 21 Continuing Calibration.......................................................................................................... 21 Lab Control Spikes (LCSs)................................................................................................... 21 Sample Duplicates............................................................................................................... 26 Surrogates........................................................................................................................... 26 E05-0210 Interim R eport#13 Tennessee R iver Page 5 of 160 3M Environm ental Labora tory E05-0210 Interim R eport#13 Field Matrix Spikes............................................................................................................... 26 Data Summary and Discussion........................................................................................................26 PFBS....................................................................................................................................26 PFHS...................................................................................................................................26 PFOS...................................................................................................................................27 PFOA[1,2 13C] Surrogate......................................................................................................27 Statistical Methods and Calculations................................................................................................ 51 Accuracy and Precision Equations........................................................................................ 51 Determination of Analytical Uncertainty................................................................................ 52 Statement of Conclusion.................................................................................................................. 52 List of Attachments.......................................................................................................................... 52 Signature Page................................................................................................................................ 53 List of Tables Table 1. Sample Results Summary................................................................................................ 10 Table 2. Study Reference Substance.............................................................................................. 15 Table 3. Study Control Substance (Surrogate).................................................................................16 Table 4. Sample Collection Information...........................................................................................16 Table 5. Instrument Parameters...................................................................................................... 17 Table 6. Liquid Chromatography Gradient Program.........................................................................17 Table 7. Mass Transitions............................................................................................................... 18 Table 8. LOQs by Extraction Set and Analysis Day..........................................................................18 Table 9. Method Blank Area Counts for Extractions Performed on 07/26/2005; Samples Analyzed on 07/26/2005 19 Table 10. Method Blank Area Counts for Extractions Performed on 07/27/2005; Samples Analyzed on 07/28/2005 and 07/30/2005............................................................................................. 20 Table 11. Method Blank Area Counts for Extraction Performed on 07/28/2005 (Centrifuged Samples); Samples Analyzed on 08/01/2005....................................................................................20 E05-0210 Interim R eport#13 Tennessee R iver Page 6 of 160 3M Environm ental Laboratory E05-0210 Interim R eport#13 Table 12. Surrogate Spiked Method Blank Recoveries..................................................................... 21 Table 13. Lab Control Spike Results: Extraction Day07/26/2005; AnalysisDay07/26/2005..............23 Table 14. Lab Control Spike Results: Extraction Day07/27/2005; AnalysisDays07/28/2005 & 07/30/2005...................................................................................................................... 24 Table 15. Lab Control Spike Results: Extraction Day 07/28/2005 (Centrifuged Samples); Analysis Day: 08/01/2005...............................................................................................................25 Table 16. PFBS Sample Results...................................................................................................... 28 Table 17. PFHS Sample Results......................................................................................................34 Table 18. PFOS Sample Results......................................................................................................40 Table 19. PFOA[1,2 13C] Surrogate Results......................................................................................46 E05-0210 Interim R eport#13 Tennessee R iver Page 7 of 160 Study Information Sponsor 3M Company Sponsor Representative Michael A. Santoro 3M Company 3M Building 0236-01-B-10 St. Paul, MN 55144 Study Director Jaisimha Kesari, P.E., DEE Weston Solutions, Inc. 1400 Weston Way West Chester, PA 19380 3M Environm ental Laboratory E05-0210 Interim R eport#13 Study Location Testing Facilities Exygen Research 3058 Research Drive State College, PA 16801 3M Environmental Laboratory 3M Building 2-3E-09 935 Bush Avenue St. Paul, MN 55144 Study Dates Study Initiation: 11/5/2004 Interim Experimental Initiation: July 26, 2005 Interim Experimental Completion: August 1, 2005 Interim Study Completion: State of Study Director's Signature Location of Archives All original raw data, protocol, and analytical report have been archived at the 3M Environmental Laboratory according to 40 CFR Part 792. The test substance and analytical reference standard reserve samples are archived at the 3M Environmental Laboratory according to 40 CFR Part 792. E05-0210 Interim R eport#13 Tennessee R iver Page 8 of 160 3M Environm ental Laboratory E05-0210 Interim R eport#13 Summary and Introduction The 3M Environmental Laboratory extracted and analyzed water samples collected from the Tennessee River by Weston Solutions personnel on July 18-22, 2005. Samples were collected from Tennessee River Mile (TRM) 307, located six miles upstream from the 3M Decatur Alabama facility to TRM 254, located 47 miles downstream from the 3M facility. Samples were submitted for analysis as part of 3M Environmental Laboratory project number E05-0210 (3M Decatur Fluorochemical GLP Monitoring Program, Interim Study #13: Analysis of Tennessee River Water Samples). Water samples were analyzed for perfluorooctane sulfonate (PFOS), perfluorohexane sulfonate (PFHS), and perfluorobutane sulfonate (PFBS) using 3M Environmental Laboratory Method ETS 8-154.1 "Determination of Perfluorinated Acids, Alcohols, Amides, and Sulfonates in Water by Solid Phase Extractions and High Performance Liquid Chromatography/Mass Spectrometry" in accordance with Exygen Research Protocol P0001131 (Attachment C). The analytical start date for this interim report was July 26, 2005 and the analytical termination date was August 1,2005. Sample containers were prepared at the 3M Environmental Laboratory. Sample containers for each sampling location included a field sample, field sample duplicate, low field spike (0.05 ng/mL) and high field spike (0.5 ng/mL). Each empty container was marked with a "fill to here" line and was fortified with a surrogate spike or an appropriate matrix spike solution containing the surrogate and the three target analytes prior to being sent to the field for sample collection. Table 1 below summarizes the sample results. The average between the sample and the sample duplicate is provided along with the relative percent difference (%RPD) if applicable. The limit of quantitation (LOQ) for all the analytes varied depending on the extraction and analysis dates. All results for quality control samples prepared and analyzed with the samples will be reported and discussed elsewhere in this report. E05-0210 Interim R eport#13 Tennessee R iver Page 9 of 160 3M Environm ental Labora tory E05-0210 Interim R eport#13 Table 1. Sample Results Summary Sample Description DLS-SW-TRM307-0-050718 DLS-SW-TRM307-0-050718 Dup DLS-SW-TRM301-0-050718 DLS-SW-TRM301-0-050718 Dup DLS-SW-TRM301-F-050718 DLS-SW-TRM301-F-050718 Dup DLS-SW-TRM295-0-050719 DLS-SW-TRM295-0-050719 Dup DLS-SW-TRM289-0-050719 DLS-SW-TRM289-0-050719 Dup DLS-SW-TRM289-F-050719 DLS-SW-TRM289-F-050719 Dup DLS-SW-TRM283-0-050719 DLS-SW-TRM283-0-050719 Dup DLS-SW-TRM277-0-050719 DLS-SW-TRM277-0-050719 Dup DLS-SW-TRM277-F-050719 DLS-SW-TRM277-F-050719 Dup DLS-SW-TRM271 -0-050719 DLS-SW-TRM271-0-050719 Dup 3M LIMS ID Number E05-0210-86358 E05-0210-86359 Average %RPD E05-0210-86362 E05-0210-86363 Average %RPD E05-0210-86366 E05-0210-86367 Average %RPD E05-0210-86372 E05-0210-86373 Average %RPD E05-0210-86374 E05-0210-86375 Average %RPD E05-0210-86378 E05-0210-86379 Average %RPD E05-0210-86382 E05-0210-86383 Average %RPD E05-0210-86386 E05-0210-86387 Average %RPD E05-0210-86390 E05-0210-86391 Average %RPD E05-0210-86394 E05-0210-86395 Average %RPD (1)PFBS (ng/mL) (1)PFHS (ng/mL) (1)PFOS (ng/mL) < 0.00221 < 0.00221 |2|< 0.00221 NA < 0.00104 < 0.00104 |2|< 0.00104 NA 0.0067 < 0.00496 |2|< 0.00496 NA < 0.00221 < 0.00221 |2|< 0.00221 NA < 0.00104 0.00134 |3|0 .0 0 1 3 4 NA < 0.00496 < 0.00496 |2|< 0.00496 NA < 0.00221 < 0.00221 |2|< 0.00221 NA < 0.00104 < 0.00104 |2|< 0.00104 NA < 0.00496 < 0.00496 |2|< 0.00496 NA |3|0.0143 |3|0.0150 0.0146 4.79 0.0115 0.0118 0.0116 2.59 0.0736 0.0747 0.0742 1.48 0.0225 0.0213 0.0218 5.50 0.0133 0.0123 0.0127 7.87 0.0763 0.0722 0.0742 5.52 0.0197 0.0201 0.0199 2.01 0.0129 0.0120 0.0124 7.26 0.0588 0.0462 0.0524 24.0 0.0203 0.0181 0.0192 11.4 0.00888 0.00883 0.00886 0.564 0.0528 0.0703 0.0616 28.4 0.0115 0.0123 0.0119 6.72 0.00579 0.00704 0.00642 19.5 0.0336 0.0530 0.0432 44.9 0.0111 0.0125 0.0118 11.9 0.00609 0.00672 0.00640 9.84 0.0304 0.0390 0.0347 24.8 0.0133 0.0142 0.0138 6.52 0.00512 0.00574 0.00543 11.4 0.0320 0.0332 0.0326 3.68 E05-0210 Interim R eport#13 Tennessee R iver Page 10 of 160 3M Environm ental Labora tory E05-0210 Interim R eport#13 Table 1. Sample Results Summary Continued. Sample Description DLS-SW-TRM265-0-050719 DLS-SW-TRM265-0-050719 Dup DLS-SW-TRM261-2-050720 DLS-SW-TRM261-2-050720 Dup DLS-SW-TRM261 -0-050720 DLS-SW-TRM261-0-050720 Dup DLS-SW-TRM261 -F-050720 DLS-SW-TRM261 -F-050720 Dup DLS-SW-TRM256-0-050720 DLS-SW-TRM256-0-050720 Dup DLS-SW-TRM254-0-050720 DLS-SW-TRM254-0-050720 Dup DLS-SW-TRM254-F-050720 DLS-SW-TRM254-F-050720 Dup DLS-SW-TRM254-2-050720 DLS-SW-TRM254-2-050720 Dup DXS-SW-T01001 -0-050721 DXS-SW-T01001-0-050721 Dup DXS-SW-T01002-0-050721 DXS-SW-T01002-0-050721 Dup 3M LIMS ID Number E05-0210-86398 E05-0210-86399 Average %RPD E05-0210-86402 E05-0210-86403 Average %RPD E05-0210-86406 E05-0210-86407 Average %RPD E05-0210-86410 E05-0210-86411 Average %RPD E05-0210-86414 E05-0210-86415 Average %RPD E05-0210-86418 E05-0210-86419 Average %RPD E05-0210-86422 E05-0210-86423 Average %RPD E05-0210-86426 E05-0210-86427 Average %RPD E05-0210-86430 E05-0210-86431 Average %RPD E05-0210-86434 E05-0210-86435 Average %RPD (1)PFBS (ng/mL) (1)PFHS (ng/mL) (1)PFOS (ng/mL) 0.0103 0.0104 0.0104 0.962 0.00525 0.00542 0.00534 3.18 0.0377 0.0375 0.0375 0.533 < 0.00221 < 0.00221 (2)< 0.00221 NA < 0.00104 < 0.00104 (2)< 0.00104 NA < 0.00496 < 0.00496 (2)< 0.00496 NA 0.0118 0.0110 0.0114 7.02 0.0052 0.00481 0.00500 7.80 0.0324 0.0330 0.0323 1.86 0.0116 0.0107 0.0112 8.04 0.00542 0.00486 0.00514 10.9 0.0333 0.0232 0.0282 35.8 0.0111 0.0125 0.0118 11.9 0.00485 0.0047 0.00478 3.14 0.0326 0.0342 0.0333 4.80 0.0124 0.0120 0.0121 3.30 0.0055 0.00481 0.00516 13.4 0.0352 0.0341 0.0346 3.18 0.0128 0.0121 0.0124 5.64 0.00533 0.00525 0.00528 1.52 0.0300 0.0342 0.0321 13.1 < 0.00221 < 0.00221 (2)<0.00221 NA < 0.00104 < 0.00104 (2)< 0 .0 0 1 0 4 NA < 0.00992 < 0.00992 (2)< 0 .0 0 9 9 2 NA 0.0215 0.0201 0.0208 6.73 0.0121 0.0116 0.0118 4.24 0.0739 0.0680 0.0710 8.31 0.0161 0.0164 0.0162 1.85 0.0101 0.00959 0.00984 5.18 0.0607 0.0594 0.0600 2.17 E05-0210 Interim R eport#13 Tennessee R iver Page 11 o f 160 3M Environm ental Labora tory E05-0210 Interim R eport#13 Table 1. Sample Results Summary Continued. Sample Description DXS-SW-T01003-0-050721 DXS-SW-T01003-0-050721 Dup DXS-SW-T01004-0-050721 DXS-SW-T01004-0-050721 Dup DXS-SW-T01005-0-050721 DXS-SW-T01005-0-050721 Dup DXS-SW-T01006-0-050721 DXS-SW-T01006-0-050721 Dup DXS-SW-T01007-0-050721 DXS-SW-T01007-0-050721 Dup DXS-SW-T01008-0-050721 DXS-SW-T01008-0-050721 Dup DXS-SW-T01009-0-050721 DXS-SW-T01009-0-050721 Dup DXS-SW-T01010-0-050721 DXS-SW-T01010-0-050721 Dup DXS-SW-T02001 -0-050721 DXS-SW-T02001-0-050721 Dup DXS-S W -T 0 200 2-0 -050 721 DXS-SW-T02002-0-050721 Dup 3M LIMS ID Number E05-0210-86438 E05-0210-86439 Average %RPD E05-0210-86442 E05-0210-86443 Average %RPD E05-0210-86446 E05-0210-86447 Average %RPD E05-0210-86450 E05-0210-86451 Average %RPD E05-0210-86454 E05-0210-86455 Average %RPD E05-0210-86458 E05-0210-86459 Average %RPD E05-0210-86462 E05-0210-86463 Average %RPD E05-0210-86466 E05-0210-86467 Average %RPD E05-0210-86470 E05-0210-86471 Average %RPD E05-0210-86474 E05-0210-86475 Average %RPD (1)PFBS (ng/mL) (1)PFHS (ng/mL) 0.0165 0.0166 0.0166 0.602 0.00827 0.00941 0.00884 12.9 0.0158 0.0165 0.0162 4.32 0.00967 0.0101 0.00988 4.35 0.00957 0.00985 0.00971 2.88 0.00633 0.00667 0.00649 5.24 0.00432 0.00366 0.00399 16.5 0.00346 0.00333 0.00340 3.82 0.00566 0.00564 0.00565 0.354 0.00419 0.00429 0.00424 2.36 0.00369 0.00502 0.00436 30.5 0.00316 0.00328 0.00321 3.74 0.00491 0.00444 0.00468 10.0 0.00301 0.0033 0.00316 9.18 0.00395 0.00385 0.00390 2.56 0.00332 0.00277 0.00304 18.1 0.0414 0.0423 0.0418 2.15 0.0205 0.0187 0.0195 9.23 0.0306 0.0316 0.0311 3.22 0.0124 0.0126 0.0125 1.60 (1)PFOS (ng/mL) 0.0542 0.0555 0.0548 2.37 0.056 0.0636 0.0598 12.7 0.0383 0.039 0.0386 1.81 0.0209 0.0219 0.0213 4.69 0.0273 0.0268 0.0270 1.85 0.0219 0.0217 0.0218 0.917 0.0202 0.0203 0.0202 0.495 0.0216 0.0219 0.0218 1.38 0.129 0.117 0.123 9.76 0.0707 0.0762 0.0734 7.49 E05-0210 Interim R eport#13 Tennessee R iver Page 12 of 160 3M Environm ental Labora tory E05-0210 Interim R eport#13 Table 1. Sample Results Summary Continued. Sample Description DXS-SW-T02003-0-050721 DXS-SW-T02003-0-050721 Dup DXS-SW-T02004-0-050721 DXS-SW-T02004-0-050721 Dup DXS-SW-T02005-0-050721 DXS-SW-T02005-0-050721 Dup DXS-SW-T02006-0-050721 DXS-SW-T02006-0-050721 Dup DXS-SW-T02007-0-050721 DXS-SW-T02007-0-050721 Dup DXS-SW-T02008-0-050721 DXS-SW-T02008-0-050721 Dup DXS-SW-T02009-0-050721 DXS-SW-T02009-0-050721 Dup DXS-SW-T02010-0-050721 DXS-SW-T02010-0-050721 Dup DXS-SW-T03001 -0-050721 DXS-SW-T03001-0-050721 Dup DXS-SW-T03002-0-050721 DXS-SW-T03002-0-050721 Dup 3M LIMS ID Number E05-0210-86478 E05-0210-86479 Average %RPD E05-0210-86482 E05-0210-86483 Average %RPD E05-0210-86486 E05-0210-86487 Average %RPD E05-0210-86490 E05-0210-86491 Average %RPD E05-0210-86494 E05-0210-86495 Average %RPD E05-0210-86498 E05-0210-86499 Average %RPD E05-0210-86502 E05-0210-86503 Average %RPD E05-0210-86506 E05-0210-86507 Average %RPD E05-0210-86510 E05-0210-86511 Average %RPD E05-0210-86514 E05-0210-86515 Average %RPD (1)PFBS (ng/mL) (1)PFHS (ng/mL) (1)PFOS (ng/mL) < 0.00221 < 0.00221 (2)<0.00221 NA < 0.00104 < 0.00104 (2)< 0 .0 0 1 0 4 NA < 0.00992 < 0.00992 (2)< 0 .0 0 9 9 2 NA < 0.00221 < 0.00221 (2)<0.00221 NA < 0.00104 < 0.00104 (2)< 0 .0 0 1 0 4 NA < 0.00992 < 0.00992 (2)< 0 .0 0 9 9 2 NA < 0.000883 < 0.00221 (2)<0.00221 NA < 0.0026 < 0.0026 (2)<0.0026 NA < 0.00744 < 0.0496 (2)<0.0496 NA < 0.000883 0.00101 (4)0.00101 NA < 0.0026 < 0.0026 (2)<0.0026 NA < 0.00744 0.00799 (4)0.00799 NA < 0.000883 < 0.000883 (2)< 0.000883 NA < 0.0026 < 0.0026 (2)< 0.0026 NA < 0.00744 < 0.00744 (2)< 0.00744 NA 0.00109 0.00115 0.00111 5.40 < 0.0026 < 0.0026 (2)< 0.0026 NA < 0.00744 < 0.00744 (2)< 0.00744 NA 0.00288 0.00244 0.00266 16.5 < 0.0026 < 0.0026 (2)< 0.0026 NA 0.00872 0.00883 0.00878 1.25 0.00628 0.00553 0.00590 12.7 0.00363 0.00342 0.00352 5.96 0.0426 0.0285 0.0356 39.6 0.818 0.854 0.836 4.31 0.176 0.185 0.180 5.00 0.782 0.807 0.794 3.15 0.00519 0.00424 0.00472 20.1 < 0.0026 < 0.0026 (2)< 0.0026 NA 0.0235 0.0169 0.0201 32.8 E05-0210 Interim R eport#13 Tennessee R iver Page 13 of 160 3M Environm ental Labora tory E05-0210 Interim R eport#13 Table 1. Sample Results Summary Continued. Sample Description 3M LIMS ID Number (1>PFBS (ng/mL) (1>PFHS (ng/mL) (1>PFOS (ng/mL) DXS-SW-T03003-0-050721 DXS-SW-T03003-0-050721 Dup E05-0210-86518 E05-0210-86519 Average %RPD < 0.000883 < 0.000883 (2)< 0.000883 NA < 0.0026 < 0.0026 (2)< 0.0026 NA 0.0111 0.0118 0.0114 6.14 DXS-SW-T03004-0-050722 DXS-SW-T03004-0-050722 Dup E05-0210-86522 E05-0210-86523 Average %RPD 0.0043 0.00106 0.00268 121 0.00756 < 0.0026 0.00756 NA 0.0226 0.0167 0.0196 30.1 DXS-SW-T03005-0-050722 DXS-SW-T03005-0-050722 Dup E05-0210-86526 E05-0210-86527 Average %RPD < 0.000883 0.00171 (3)0.00171 NA < 0.0026 0.00331 (4)0.00331 NA < 0.00744 0.0773 (4)0.0773 NA DXS-SW-T03006-0-050722 DXS-SW-T03006-0-050722 Dup E05-0210-86530 E05-0210-86531 Average %RPD < 0.000883 < 0.000883 (2)< 0.000883 NA < 0.0026 < 0.0026 (2)< 0.0026 NA 0.00763 < 0.00744 (4)0.00763 NA DXS-SW-T03007-0-050722 DXS-SW-T03007-0-050722 Dup E05-0210-86534 E05-0210-86535 Average %RPD < 0.000883 0.00107 (4)0.00107 NA < 0.0026 0.00366 (4)0.00366 NA < 0.00744 < 0.0496 (2)<0.0496 NA DXS-SW-T03008-0-050722 DXS-SW-T03008-0-050722 Dup E05-0210-86538 E05-0210-86539 Average %RPD < 0.000883 < 0.000883 (2)< 0.000883 NA < 0.0026 < 0.0026 (2)< 0.0026 NA < 0.00744 < 0.00744 (2)< 0.00744 NA DXS-SW-T03009-0-050722 DXS-SW-T03009-0-050722 Dup E05-0210-86542 E05-0210-86543 Average %RPD < 0.000883 < 0.000883 (2)< 0.000883 NA < 0.0026 < 0.0026 (2)< 0.0026 NA 0.00809 < 0.00744 (4)0.00809 NA DXS-SW-T03010-0-050722 DXS-SW-T03010-0-050722 Dup E05-0210-86546 E05-0210-86547 0.000908 < 0.000883 < 0.0026 < 0.0026 0.00749 < 0.00744 Average (4)0.000908 (2)< 0.0026 (4)0.00749 %RPD NA NA NA (1) All results and calculations are presented with three significant figures. Sample concentrations, averages, and %RPD values may vary slightly from the raw data. The analytical uncertainty for the PFBS, PFHS, and PFOS results is 10010.6%, 10015.2%, and 10013.8%, respectively. (2) NA: Not applicable. %RPD values not determined when concentrations for the sample and/or sample duplicate are below the stated LOQ. (3) Sample descriptions were misidentified when checked into LIMS. Samples with LIMS ID numbers 86370 and 86371 were in bottles labeled low and high spike, respectively. Bottles with LIMS ID numbers 86372 and 86373 were surrogate spiked only (sample and sample duplicate bottles). (4) The average value listed is the concentration for the sample or sample duplicate that produced a value above the LOQ. A true average and %RPD between the sample and sample duplicate was not determined. E05-0210 Interim R eport#13 Tennessee R iver Page 14 of 160 3M Environm ental Labora tory E05-0210 Interim R eport#13 Test Samples Water samples (3M LIMS ID #86358-86558) were received on July 23, 2005 from Charles Young of Weston Solutions, Inc. Two hundred one (201) containers representing 48 samples and an equipment rinse blank were submitted for analysis. The samples were logged in by 3M Environmental Laboratory professional services personnel and placed in refrigerated storage prior to extraction on July 26-28, 2005. Reference Substances Table 2 lists the pertinent information regarding the reference substance used for this study. Table 2. Study Reference Substance. Reference Substance PFBS PFHS PFOS Chemical Name Perfluorobutanesulfonate Perfluorohexanesulfonate Perfluorooctanesulfonate Chemical Formula Identifier (1)S o u rc e Expiration Date Storage Conditions Chemical Lot Number TCR Number C4FgSOsK+ CAS # 29420-49-3 3M 10/17/2006 Frozen 101 TCR-116 (99030-023) C6F1 3 SO3 K CAS # 3871-99-6 3M 10/18/2006 Frozen NB# 120067-69 TCR-83 (SE036) C8F17SO3'K CAS # 2795-39-3 3M 8/31/2006 Frozen 171 TCR-696 Physical Description W hite Powder White Powder White powder Purity (2)Solubility (1) (2) 96.7% 98.6% 86.4% 54,400 ppm No Information available Documentation regarding synthesis of the test substances is located at the source. 680 ppm All test substances are believed to be soluble in water at the levels to be investigated. No visual precipitates observed. Control Substance PFOA [1,2 13C] was analyzed as a surrogate against a multi-level extracted calibration curve. A known amount of PFOA [1,2 13C] was spiked into each sample container in the laboratory prior to sample collection. Table 3 lists the pertinent information regarding the study control substance. E05-0210 Interim R eport#13 Tennessee R iver Page 15 of 160 3M Environm ental Labora tory E05-0210 Interim R eport#13 Table 3. Study Control Substance (Surrogate). Reference Substance Chemical Name Chemical Formula Identifier Source Expiration Date Storage Conditions Chemical Lot Number TCR Number Physical Description Purity PFOA [1,2 13C] Perfluorooctanoic Acid C6 F1 5 [1 3 C ]F2 [1 3 C]OOH N/A Perkin Elmer 03/29/2009 Frozen 3507-195 TCR-744 W hite powder >97% Method Summaries Sample Collection Samples were collected on July 18-23, 2005 in pre-rinsed NalgeneTM (low-density polyethylene) bottles prepared at the 3M Environmental Laboratory on July 14, 2005. Prior to sample collection, all bottles were spiked in the laboratory with a known volume of a surrogate solution or appropriate matrix spiking solution containing the surrogate and the analyte of interest. Four collection bottles were associated with each individual sampling location: sample, sample duplicate, low level field matrix spike (LS), and high level field matrix spike (HS). Additionally, three "Trip Blank" sample sets were prepared. A Trip Blank set consisted of three individual bottles: trip blank sample, trip blank low spike, and trip blank high spike. The surrogate and matrix spike levels for the trip blanks were the same as the samples. Table 4 below details the spike amounts for the four bottles comprising the sample location set. Table 4. Sample Collection Information (1)Bottle Description Nominal Final Volume Collected (mL) PFBS Final Spike Concentration (ng/mL) PFHS PFOS PFOA [1 ,2 13C] (Surrogate) Sample 450 NA NA NA Sample Dup 450 NA NA NA Low Field Matrix Spike (LS) 450 0.0441 0.0521 0.0496 High Field Matrix Spike (HS) 450 0.441 0.521 0.496 (1) The sample description was assigned in the field by Weston Solutions personnel. 0.274 0.274 0.254 0.254 E05-0210 Interim R eport#13 Tennessee R iver Page 16 of 160 3M Environm ental Labora tory E05-0210 Interim R eport#13 Preparatory and Analytical Methods Extraction All samples, calibration standards, and associated quality control samples were extracted using a modified procedure of ETS-8-154.1 "Determination of Perfluorinated Acids, Alcohols, Amides, and Sulfonates in Water by Solid Phase Extractions and High Performance Liquid Chromatography/Mass Spectrometry". Briefly, approximately 100 mL of sample were loaded onto a pre-conditioned Waters C18 solid phase extraction (SPE) cartridge (Sep-Pak, 6 cc) using a vacuum manifold. The loaded cartridges were then eluted with 2 mL of methanol This extraction procedure concentrates the samples by a factor of fifty. (Initial volume = 100 mL, final volume = 2 mL). As written, ETS-8-154.1 calls for a 40 mL and 5 mL extraction and elution volume, respectively. Due to the large number of samples associated with this project, samples were extracted over the course of three days (July 26-July 28, 2005). The extracted calibration curve used to quantify the data was only prepared on the first extraction day; however, method blanks and lab control spikes were prepared each day samples were extracted. During the second day of extraction, six samples clogged the SPE cartridge preventing complete loading of the sample. On the third extraction day, these six samples were centrifuged at 6000 rcf (relative centrifugal force) for 20 minutes prior to loading the SPE cartridge. Method blanks and lab control spikes were also centrifuged and extracted in a similar manner. Analysis All sample and quality control extracts were analyzed for PFBS, PFHS, PFOS, and PFOA [1,2 13C] using high performance liquid chromatography/ tandem mass spectrometry (HPLC/MS/MS). Pertinent instrument parameters, the liquid chromatography gradient program, and the specific mass transitions analyzed are described in the tables below. Table 5. Instrument Parameters Instrument Name Liquid Chromatograph Guard column Analytical column Injection Volume Mass Spectrometer Electrode Ion Source Polarity Software ETStan Agilent 1100 Betasil C18 2X50, 5 urn Betasil C18 (2.1 mm X 100 mm), 5 ixm 5 ixL Applied Biosystems API 4000 Z-spray Turbo Spray Negative Analyst 1.4.1 Table 6. Liquid Chromatography Gradient Program Step Number 0 1 2 3 4 5 Total Time (min) 0 1.0 14.5 15.5 16.5 20.0 Flow Rate (pL/min) 300 300 300 300 300 300 Percent A (2 m M ammonium acetate) 80.0 80.0 10.0 10.0 80.0 80.0 Percent B (Methanol) 20.0 20.0 90.0 90.0 20.0 20.0 E05-0210 Interim R eport#13 Tennessee R iver Page 17 of 160 3M Environm ental Laboratory E05-0210 Interim R eport#13 Table 7. Mass Transitions Analyte (1)PFBS (1)PFHS (1)PFOS PFOA [1,2 13C] Mass Transition Q1/Q3 299/80 299/99 399/80 399/99 499/130 499/99 499/80 415/370 Dwell Time (msec) 150 150 150 150 150 150 150 150 (1) The individual transitions were summed to produce a "total ion chromatogram" (TIC). The TIC was used for quantitation. A nalytical Results Calibration Calibration standards were prepared by spiking known amounts of stock solutions containing PFBS, PFHS, PFOS, and PFOA [1,2 13C] into 100 mL of ASTM type I water. Each spiked water standard was then extracted in the same manner as the collected samples. A total of twelve spiked standards ranging from 0.001 ng/mL to 2.5 ng/mL (nominal) were prepared. A quadratic, 1/x weighted, calibration curve was used to fit the data for each analyte. The data was not forced through zero during the fitting process. Calculating the standard concentration using the peak area counts and the resultant calibration curve confirmed accuracy of each curve point. Each extracted calibration standard used to generate the final calibration curve met the method calibration accuracy requirement of 10025%. Coefficients of determination (r2) were greater than 0.995 for all analytes. Limit of Quantitation (LOQ) The LOQ for this analysis, as defined in ETS-8-154.1, is the lowest non-zero calibration standard in the curve in which the area counts are at least twice those of the method blank(s). The LOQ varied from day to day depending on instrument sensitivity and method blank area counts for the extraction set. This will be presented and discussed in more detail in the section below regarding method blanks. Table 8 below provides the individual LOQs for the three analytes of interest. Table 8. LOQs by Extraction Set and Analysis Day Extraction Set 07/26/2005 07/27/2005 07/28/2005 (Centrifuged Samples) Analysis Day 07/26/2005 07/28/2005 07/30/2005 08/01/2005 LOQ PFBS (ng/mL) PFHS (ng/mL) PFOS (ng/mL) 0.00221 0.00221 0.000883 0.00221 0.00104 0.00104 0.00260 0.00260 0.00496 0.00992 0.00744 0.0496 E05-0210 Interim R eport#13 Tennessee R iver Page 18 of 160 3M Environm ental Laboratory E05-0210 Interim R eport#13 Blanks Three types of blanks were prepared and analyzed with the samples: method blanks, solvent blanks, and field/trip blanks. Each blank type is described below. Solvent Blank Several methanol solvent blanks were analyzed to assess system contamination and/or instrument carryover. Analyte peak area counts in all blank samples were less than half the area counts of the calibration standard used to establish the LOQ. Method Blank Over the three days in which samples, calibration standards, and quality control samples were extracted, several method blanks were prepared by loading 100 mL of ASTM Type I water onto a C18 SPE cartridge and eluting with 2 mL of methanol using the same extraction procedure as the samples. Method blanks were prepared to evaluate the levels of background contamination in the overall extraction process (glassware, SPE cartridges, etc.) Approximately half of the method blanks prepared each day were spiked with a known amount of PFOA [1,2 13C] surrogate prior to extraction. Table 9 and Table 10 list the analyte area counts for the method blanks and the corresponding LOQ standard for the given analysis day. Table 11 lists the analyte area counts of the method blanks associated with the centrifuged samples. The PFOS LOQ for these six samples was raised to 0.0496 ng/mL due to higher amounts of PFOS detected in the method blanks prepared on that day. Surrogate recoveries for the spiked method blanks are presented in Table 12. Method blanks spiked with surrogate solution produced an average recovery of 176% with a RSD of 8.45%. An older spiking solution that had been used extensively since preparation is hypothesized to be the underlying cause of the high recovery. Solvent evaporation due to opening the storage container several times is plausible. Table 9. Method Blank Area Counts for Extractions Performed on 07/26/2005; Samples Analyzed on 07/26/2005. Sample Name Sample Comment s050726a028 Method Blank-1 s050726a029 Method Blank-2 s050726a030 Method Blank-3 s050726a031 Method Blank-4 s050726a032 Method Blank-5 s050726a033 Method Blank-6 s050726a034 Method Blank-7 s050726a035 Method Blank-8 LOQ Standard Area Counts LOQ Standard Concentration Sample ID MB-050726-1 MB-050726-2 MB-050726-3 MB-050726-4 MB-050726-5 MB-050726-6 MB-050726-7 MB-050726-8 PFBS Area PFHS Area PFOS Area Counts Counts Counts 2250 2770 4317 2314 8191 3771 3668 2600 26255 0.00221 1301 1881 1793 1754 1642 1804 1546 949 8250 0.00104 3358 5306 5128 2802 4514 4259 4700 3662 12939 0.00496 E05-0210 Interim R eport#13 Tennessee R iver Page 19 of 160 3M Environm ental Laboratory E05-0210 Interim R eport#13 Table 10. Method Blank Area Counts for Extractions Performed on 07/27/2005; Samples Analyzed on 07/28/2005 and 07/30/2005. Sample Name Sample Comment s050728a025 s050728a026 s050728a027 s050728a028 Method Blank-1 Method Blank-2 Method Blank-5 Method Blank-6 LOQ Standard Area Counts LOQ Standard Concentration s050730a025 Method Blank-3 s050730a026 Method Blank-4 s050730a027 Method Blank-7 s050730a028 Method Blank-8 LOQ Standard Area Counts LOQ Standard Concentration Sample ID MB-050727-1 MB-050727-2 MB-050727-5 MB-050727-6 MB-050727-3 MB-050727-4 MB-050727-7 MB-050727-8 PFBS Area PFHS Area PFOS Area Counts Counts Counts 3215 4573 3369 3316 1001 1930 39 0 3488 9299 3045 4497 27360 0.00221 3414 5089 3632 3941 14056 0.000880 6707 0.00104 28 213 48 1796 15862 0.00260 21214 0.00992 4043 9052 5865 9632 20564 0.00744 Table 11. Method Blank Area Counts for Extraction Performed on 07/28/2005 (Centrifuged Samples); Samples Analyzed on 08/01/2005. Sample Name Sample Comment Sample ID PFBS Area PFHS Area PFOS Area Counts Counts Counts Centrifuge s050801a025 Method Blank-1 MB-050728-1 7312 3369 27461 Yes s050801a026 Method Blank-2 MB-050728-2 3505 2182 24740 Yes s050801a027 Method Blank-3 MB-050728-3 7853 5636 42506 No s050801a028 Method Blank-4 MB-050728-4 4363 2456 28031 No s050801a029 Method Blank-5 MB-050728-5 2528 925 19878 Yes s050801a030 Method Blank-6 MB-050728-6 2673 1104 18783 Yes s050801a031 Method Blank-7 MB-050728-7 (1)73234 (1)105441 (1)1495463 No s050801a032 Method Blank-8 MB-050728-8 (1)29997 (1)38166 (1)237882 No LOQ Standard Area Counts 26908 15530 107559 LOQ Standard Concentration 0.00221 0.00260 0.0496 (1) The area counts for Method Blank-7 and Method Blank-8 were determined to be statistical outliers using Dixon's Q test. These data points were excluded when determining the LOQs for the centrifuged samples. E05-0210 Interim R eport#13 Tennessee R iver Page 20 of 160 3M Environm ental Laboratory E05-0210 Interim R eport#13 Table 12. Surrogate Spiked Method Blank Recoveries. Sample Name Sample Comment Sample ID Analyte Concentration (ng/mL) Calculated Concentration PFOA [ 1 ,2 13C] ng/mL Percent Surrogate Recovery s050726a032 Method Blank-5 MB-050726-5 0.102 0.197 193 s050726a033 Method Blank-6 MB-050726-6 0.102 0.169 166 s050726a034 Method Blank-7 MB-050726-7 0.102 0.197 193 s050726a035 Method Blank-8 MB-050726-8 0.102 0.175 172 s050728a027 Method Blank-5 MB-050727-5 0.102 0.16 157 s050728a028 Method Blank-6 MB-050727-6 0.102 0.163 160 s050730a027 Method Blank-7 MB-050727-7 0.102 0.166 163 s050730a028 Method Blank-8 MB-050727-8 0.102 <0.00249 (1)NA s050801a029 Method Blank-5 MB-050728-5 0.102 0.184 180 s050801a030 Method Blank-6 MB-050728-6 0.102 0.178 174 s050801a031 Method Blank-7 MB-050728-7 0.102 0.206 202 s050801a032 Method Blank-8 MB-050728-8 0.102 0.182 178 (1) NA: Not applicable. Surrogate not measured in final extract suggesting that the solution was inadvertently not spiked prior to extraction. (2) The average surrogate recovery in spiked method blanks was 176% (RSD=8.45%). Trip Blank Prior to sample collection, three sample containers were filled with 450 mL of ASTM Type I water, spiked with surrogate, sealed, and shipped to the sample collection site along with the empty containers. These samples were analyzed as field/trip blanks. Trip blanks serve as additional method blanks that account for any storage conditions and/or holding time issues that the samples may experience. The resultant field/trip blank concentrations for PFBS, PFHS, and PFOS were <0.000883 ng/mL, <0.00260 ng/mL, and <0.00744 ng/mL, respectively. System Suitability A 1.0 ng/mL (nominal concentration) extracted calibration standard was analyzed in triplicate at the beginning and end of the analytical sequence to demonstrate overall system suitability. For all analytes, the relative standard deviation (RSD) of the analyte peak area counts and the RSD of the peak retention times was less than 5% and 2%, respectively, which met method criteria. Continuing Calibration During the course of the analytical sequence, several continuing calibration verification samples (CCVs) were analyzed to confirm that the instrument response and the initial calibration curve was still in control. All CCVs for all four analytes produced recoveries within 25%, which met method criteria. Lab Control Spikes (LCSs) Replicate low (0.025 ng/mL nominal concentration) and high (1.25 ng/mL nominal concentration) lab control spikes were prepared and extracted each day samples or calibration standards were extracted. LCSs were prepared by spiking known amounts of the analytes into 100 mL of ASTM Type I water to produce the desired concentration. The spiked water samples were extracted and analyzed in the same manner as the samples. Individual LCS results, along with the average and percent RSD, for each extraction day are E05-0210 Interim R eport#13 Tennessee R iver Page 21 of 160 3M Environm ental Laboratory E05-0210 Interim R eport#13 presented in the data tables below. LCSs were used to evaluate method accuracy and precision which was then used to determine analytical uncertainty for each of the analytes reported with the results in Table 1. Information on how the analytical uncertainty was determined is provided at the end of this report in the "Statistical Methods and Calculations" section. For the third extraction day (07/28/2005 - centrifuged samples), a total of twelve LCSs were prepared. Six of the twelve replicates (three low, three high) were centrifuged before extraction in a manner similar to the samples. The remaining six replicates (three low, three high) were not centrifuged prior to extraction. These samples were prepared to verify that the added centrifugation step did not result in loss of target analytes. All LCS recoveries for the first two extraction days met method criteria for accuracy (percent recovery:10025%) and precision (%RSD < 15%) for all analytes (Table 13 and Table 14). However, for the third extraction day, three LCS measurements exceeded method criteria for accuracy (two high level replicates of PFOA[1,2 C] both with 131% recovery and one high level PFBS with 129% recovery). Despite these three exceedences, the LCS recoveries demonstrate excellent accuracy and precision, at both individual levels, and when considered collectively for this extraction day. Therefore, the samples extracted on that day are considered to be accurate within the stated method uncertainty. The overall project accuracy (percent recovery) and precision (percent RSD) was determined for each analyte using the LCS data from the three separate extraction days. The overall project accuracy and precision for PFBS, PFHS, and PFOS was 1073.49%, 1104.72%, and 1085.79%, respectively. These values were used to estimate the analytical uncertainty for each analyte which is described in detail in the Statistical Methods and Calculations section of this report. E05-0210 Interim R eport#13 Tennessee R iver Page 22 of 160 3M Environm ental Laboratory E05-0210 Interim R eport#13 Table 13. Lab Control Spike Results: Extraction Day 07/26/2005; Analysis Day 07/26/2005. Sample Description Sample ID LCS Low-1 LCS-050726-1 LCS Low-2 LCS-050726-2 LCS Low-3 LCS-050726-3 Average Low Recovery %RSD (Extraction Day 7/26/2005) LCS High-1 LCS-050726-4 LCS High-2 LCS-050726-5 LCS High-3 LCS-050726-6 Average High Recovery %RSD (Extraction Day 7/26/2005)______ Average All LCSs%RSD (Extraction Day 7/26/2005) (1) PFBS (1)PFHS VPFOS PFOA [1,2 13C] Analyte Conc. (ng/mL) 0.0221 0.0221 0.0221 Calc. Analyte Conc. Percent Conc. (ng/mL) Recovery (ng/mL) 0.0243 0.0233 0.0246 110 105 111 0.026 0.026 0.026 Calc. Analyte Conc. Percent Conc. (ng/mL) Recovery (ng/mL) 0.0285 110 0.0248 0.0298 115 0.0248 0.0294 113 0.0248 Calc. Analyte Conc. Percent Conc. (ng/mL) Recovery (ng/mL) 0.0244 0.0279 0.03 98.4 112 121 0.0249 0.0249 0.0249 Calc. Conc. Percent (ng/mL) Recovery 0.0262 0.0283 0.0275 105 114 110 109% 2.83% 112% 2.28% 111% 10.3% 110% 4.26% 1.10 1.15 104 1.30 1.4 108 1.24 1.35 109 1.25 1.25 100 1.10 1.18 107 1.30 1.41 108 1.24 1.35 109 1.25 1.18 94.4 1.10 1.19 108 1.30 1.44 111 1.24 1.42 114 1.25 1.23 98.4 107%1.77% 109%1.47% 111%2.94% 97.6% 2.96% 108%2.40% 111%2.43% 111%6.78% 104% 7.15% (1). Table displays rounded values for all concentration and percent recovery values (3 significant figures). Recovery and RSD values may vary slightly from the values in the raw data. E05-0210 Interim R eport#13 Tennessee R iver Page 23 of 160 3M Environm ental Laboratory E05-0210 Interim R eport#13 Table 14. Lab Control Spike Results: Extraction Day 07/27/2005; Analysis Days 07/28/2005 & 07/30/2005. "PFBS PFHS 1PFOS (1)PFOA [1,2 13C] Sample Description Sample ID Analyte Conc. (ng/mL) Calc. Analyte Conc. Percent Conc. (ng/mL) Recovery (ng/mL) Calc. Analyte Conc. Percent Conc. (ng/mL) Recovery (ng/mL) Calc. Analyte Conc. Percent Conc. (ng/mL) Recovery (ng/mL) Calc. Conc. Percent (ng/mL) Recovery LCS Low-1 LCS-050727-1 0.0221 0.0238 108 0.026 0.029 112 0.0248 0.0243 98.0 0.0249 0.0255 102 LCS Low-2 LCS-050727-2 0.0221 0.0238 108 0.026 0.0286 110 0.0248 0.0241 97.2 0.0249 0.0275 110 LCS Low-3 LCS-050727-3 0.0221 0.0233 105 0.026 0.0278 107 0.0248 0.0257 104 0.0249 0.0241 96.8 Average Low Recovery %RSD (Extraction Day 7/27/2005)_____ 107% 1.22% 109% 2.15% 99.6%3.53% 103% 6.86% LCS High-1 LCS-050727-4 1.1 1.11 101 1.3 1.37 105 1.24 1.31 106 1.25 1.08 86.4 LCS High-2 LCS-050727-5 1.1 1.14 104 1.3 1.41 108 1.24 1.32 106 1.25 1.02 81.6 LCS High-3 LCS-050727-6 1.1 1.12 102 1.3 1.37 105 1.24 1.28 103 1.25 1.05 84.0 Average High Recovery %RSD (Extraction Day 7/27/2005)______ 102%1.36% 106%1.67% 105%1.60% 84.0%2.86% Average All LCSs%RSD (Extraction Day 7/27/2005) 104%2.78% 108%2.33% 102%3.81% 93.6%12.3% (1). Table displays rounded values for all concentration and percent recovery values (3 significant figures). Recovery and RSD values may vary slightly from the values in the raw data. E05-0210 Interim R eport#13 Tennessee R iver Page 24 of 160 3M Environm ental Laboratory E05-0210 Interim R eport#13 Table 15. Lab Control Spike Results: Extraction Day 07/28/2005 (Centrifuged Samples); Analysis Day: 08/01/2005. (1)PFBS (1)PFHS (1)PFOS (1)PFOA [1,2 13C] Sample Description Sample ID Analyte Calculated Analyte Calculated Analyte Calculated Analyte Calculated Conc. Conc. Percent Conc. Conc. Percent Conc. Conc. Percent Conc. Conc. Percent (ng/mL) (ng/mL) Recovery (ng/mL) (ng/mL) Recovery (ng/mL) (ng/mL) Recovery (ng/mL) (ng/mL) Recovery LCS Low-1 Centrifuged LCS-050728-1 LCS Low-2 Centrifuged LCS-050728-2 LCS Low-3 Centrifuged LCS-050728-3 Average Low Centrifuged (Extraction Day 7/28/2005) LCS High-1 Centrifuged LCS-050728-4 LCS High-2 Centrifuged LCS-050728-5 LCS High-3 Centrifuged LCS-050728-6 Average High Centrifuged (Extraction Day 7/28/2005) Average A ll Centrifuged LCSs (Extraction Day 7/28/2005) LCS Low-7 LCS-050728-7 LCS Low- 8 LCS-050728-8 LCS Low-9 LCS-050728-9 Average Low Non-Centrifugeed (Extraction Day 7/28/2005) LCS High-10 LCS-050728-10 LCS High-11 LCS-050728-11 LCS High-12 LCS-050728-12 Average High Non-Centrifuged (Extraction Day 7/28/2005) Average All Non-Centrifuged LCSs (Extraction Day 7/28/2005) 0.0221 0.0221 0.0221 1.1 1.1 1.1 0.0221 0.0221 0.0221 1.1 1.1 1.1 0.024 0.0241 0.0235 108%1.35% 1.2 1.2 1.16 108%1.95% 108%1.50% 0.0236 0.0263 0.023 110%7.23 1 .1 2 1.17 1.15 104%2.19% 107%5.69% 108 109 106 109 109 105 107 119 104 102 106 104 0.026 0.026 0.026 0.029 0.0284 0.0288 112 109 111 110%1.06% 1.3 1.43 1 1 0 1.3 1.38 106 1.3 1.37 105 107%2.47% 109%2.31% 0.026 0.026 0.026 0.0294 0.0335 0.0304 113 129 117 120%6.87% 1.3 1.36 105 1.3 1.36 105 1.3 1.38 106 105% 0.888% 112%8.46% 0.0248 0.0248 0.0248 (2)<0.0496 (2)<0.0496 ((2)<0.0496 NA NA NA NA 1.24 1.24 1.24 1.42 1.36 1.35 114 110 109 111%2.75% (3) 111%2.75% 0.0248 0.0248 0.0248 (2)<0.0496 (2)<0.0496 ((2)<0.0496 NA NA NA NA 1.24 1.29 1.24 1.36 1.24 1.37 108% 3.25% 104 110 110 (3) 108%3.25% 0.0249 0.0249 0.0249 0.028 0.0307 0.0297 112 123 119 118%5.48% 1.25 1.45 116 1.25 1.49 119 1.25 1.45 116 117%1.58% 118%3.16% 0.0249 0.0249 0.0249 0.0295 0.0326 0.0289 118 131 116 121%7.98% 1.25 1.43 1.25 1.45 1.25 1.64 114 116 131 120%7.69% 121%6.41% (1) . Table displays rounded values for all concentration and percent recovery values (3 significant figures). Recovery and RSD values may vary slightly from the values in the raw data. (2) The PFOS LOQ for this analysis set was raised to 0.0496 due to method blank contamination. LCS results and recoveries were not reported. (3) The average value and %RSD were calculated using only the high level results. E05-0210 Interim R eport#13 Tennessee R iver Page 25 of 160 3M Environm ental Laboratory E05-0210 Interim R eport#13 Sample Duplicates Because a field sample duplicate (separate container) was collected at each sampling location, duplicate/replicate extractions of a given sample were not performed. Overall method precision was determined using surrogate spikes and LCSs. Surrogates Although not specified in the ETS 8-154.1, PFOA [1,2 13C] was added to all samples and sample spikes as a surrogate to evaluate overall method performance. The final PFOA[1,213C] concentration was 0.274 ng/mL. Surrogate recoveries are reported in the next section with sample data and will be discussed in further detail in the Data Summary and Discussion section below. Field Matrix Spikes Low (nominal concentration of 0.05 ng/mL) and high (nominal concentration 0.5 ng/mL) field matrix spikes were collected at each sampling point to verify that the analytical method is applicable to the collected matrix. The PFOA [1,2 13C] surrogate was added to each field matrix spike at a final concentration of 0.254 ng/mL. Field matrix spike recoveries within 10030% confirm that "unknown" components in the sample matrix do not interfere with the extraction and analysis of the analytes of interest. Field matrix spikes will be presented in the next section with the sample data. Data Summary and Discussion The data tables below individually summarize the sample results and field matrix spike recoveries for each of the three analytes and the surrogate for all of the sampling locations plus the three trip blanks. PFBS PFBS matrix spike recoveries were within method acceptance criteria of 10030% for all sampling locations with the following exceptions: 3M LIMS sample numbers 86448 (low spike 174%), 86452 (low spike 137%), 86456 (low spike 223%), 86460 (low spike 134%), 86489 (high spike 136%), 86528 (low spike 133%), 86533 (high spike 134%) and 86558 (trip blank #3 high spike 18.0%). (See Table 16). For each of these instances, the corresponding high or low spike for the given location produced a spike recovery within method criteria of 10030%. Therefore, the sample results for these locations are reported within the stated method analytical uncertainty and are considered accurate to within 10030% for the given location. PFHS PFHS matrix spike recoveries were within method acceptance criteria of 10030% for all sampling locations with the following exceptions: 3M LIMS sample numbers 86448 (low spike 170%), 86452 (low spike 134%), 86509 (high spike 67.8%), 86512 (low spike 133%), and 86532 (low spike 54.7%). Refer to Table 17 for more information. For each of these instances, the corresponding high or low spike for the given sampling location produced a spike recovery within method criteria of 10030%. Therefore, the sample results for these locations are reported within the stated method analytical uncertainty and are considered accurate to within 10030% for the given location. E05-0210 Interim R eport#13 Tennessee R iver Page 26 of 160 3M Environm ental Laboratory E05-0210 Interim R eport#13 PFO S In general, PFOS matrix spike recoveries were within method acceptance criteria of 10030% with a few exceptions. Six locations produced recoveries that exceeded method critera for the low-level spike but were acceptable for the high-level spike. The 3M LIMS sample numbers for these instances were 86380 (low spike 136%), 86448 (low spike 191%), 86452 (low spike 139%), 86460 (low spike 136%), 86489 (low spike 138%), and 86536 (low spike 134%). With the exception of sample number 86448, the low-level spike recovery was high, but less than 150%. Because the high-level spike recoveries for these samples met method criteria, the sample results for these locations are reported within the stated method analytical accuracy and are considered accurate to within 10030%. Additionally, three sampling locations produced matrix spike recoveries outside the 10030% criteria for both spike levels: DXS-SW-T02006-0-050721 (low spike 131%, high spike 133%), DXS-SW-T02010-0-050721 (low spike 47.4%, high spike 51.9%), and DXSSW-T03006-0-050722 (low spike 2.91%, high spike 132%). For locations DXS-SW-T020060-050721 and DXS-SW-T02010-050721, the accuracy for the sample/sample duplicate results has been increased to 10050% to account for the slightly high recoveries for DXSSW-T02006-0-050721 (132% average) and the low recoveries for DXS-SW-T02010-050721 (approximately 50% average). Furthermore, the sample/sample duplicate %RPD for DXSSW-T02010-050721 was greater than 30%. The accuracy for location DXS-SW-T03006-0050722 has been increased to of 10050% based solely on the high spike recovery (132%). All sample locations with adjusted accuracy statements for PFOS have been duly footnoted in Table 18. P F O A [ 1 , 2 13C ] S u r r o g a t e Recoveries of the PFOA[1,213C] surrogate in the sample, sample duplicate, and field matrix spikes for each sampling location is provided in Table 19. When the results in Table 19 are considered collectively, one of the most noticeable observations was that the surrogate recovery was consistently low for all samples, sample duplicates, low matrix spikes and high matrix spikes. The overall average surrogate recovery was 56.1% with a RSD of 11.6%. The spiking solutions used to prepare the collection bottle spikes were evaluated prior to bottle preparation under non-GLP provisions. Surrogate recoveries from this study do not suggest that a solution preparation/calculation error is responsible for the recoveries observed here. At this time, the 3M Environmental Laboratory cannot explain the precise, but low surrogate recoveries. A separate investigation will be conducted to explore possible loss mechanisms/holding time issues. The average surrogate recovery was used as a correction factor to normalize surrogate recoveries for a potential holding time effect. Both the corrected and uncorrected recoveries are listed in Table 19. It should be emphasized that sample results for PFBS, PFHS, and PFOS were NOT corrected for surrogate recoveries. Of the 201 bottles extracted, only three produced corrected surrogate less than 70% (sample numbers 86509, 86527, and 86532). Sample number 86509 was a high-level matrix spike and exhibited low recoveries for the three target analytes (PFBS 76.7%, PFHS 67.8%, and PFOS 51.9%). Thus, the low recovery may be attributable to an error during spiking of the collection bottle. E05-0210 Interim R eport#13 Tennessee R iver Page 27 of 160 3M Environm ental Labora tory E05-0210 Interim R eport#13 Table 16. PFBS Sample Results. 3M Sample Number Sample Description E05-0210-86358 DLS-SW-TRM307-0-050718 E05-0210-86359 DLS-SW-TRM307-0-050718 Dup E05-0210-86360 DLS-SW-TRM307-0-050718 LS E05-0210-86361 DLS-SW-TRM307-0-050718 HS (1)PFBS Conc. (ng/mL) < 0.00221 < 0.00221 0.0490 0.508 E05-0210-86362 DLS-SW-TRM301-0-050718 E05-0210-86363 DLS-SW-TRM301-0-050718 Dup E05-0210-86364 DLS-SW-TRM301-0-050718 LS E05-0210-86365 DLS-SW-TRM301-0-050718 HS < 0.00221 < 0.00221 0.0465 0.533 E05-0210-86366 DLS-SW-TRM301-F-050718 E05-0210-86367 DLS-SW-TRM301-F-050718 Dup E05-0210-86368 DLS-SW-TRM301-F-050718 LS E05-0210-86369 DLS-SW-TRM301-F-050718 HS < 0.00221 < 0.00221 0.0479 0.524 E05-0210-86370 DLS-SW-TRM295-0-050719(4) E05-0210-86371 DLS-SW-TRM295-0-050719 Dup(4) E05-0210-86372 DLS-SW-TRM295-0-050719 LS(4) E05-0210-86373 DLS-SW-TRM295-0-050719 HS(4) 0.0679 0.493 0.0143 0.0150 E05-0210-86374 DLS-SW-TRM289-0-050719 E05-0210-86375 DLS-SW-TRM289-0-050719 Dup E05-0210-86376 DLS-SW-TRM289-0-050719 LS E05-0210-86377 DLS-SW-TRM289-0-050719 HS 0.0225 0.0213 0.0706 0.453 E05-0210-86378 DLS-SW-TRM289-F-050719 E05-0210-86379 DLS-SW-TRM289-F-050719 Dup E05-0210-86380 DLS-SW-TRM289-F-050719 LS E05-0210-86381 DLS-SW-TRM289-F-050719 HS 0.0197 0.0201 0.0693 0.522 E05-0210-86382 DLS-SW-TRM283-0-050719 E05-0210-86383 DLS-SW-TRM283-0-050719 Dup E05-0210-86384 DLS-SW-TRM283-0-050719 LS E05-0210-86385 DLS-SW-TRM283-0-050719 HS 0.0203 0.0181 0.0677 0.508 E05-0210-86386 DLS-SW-TRM277-0-050719 E05-0210-86387 DLS-SW-TRM277-0-050719 Dup E05-0210-86388 DLS-SW-TRM277-0-050719 LS E05-0210-86389 DLS-SW-TRM277-0-050719 HS 0.0115 0.0123 0.0607 0.515 E05-0210-86390 DLS-SW-TRM277-F-050719 E05-0210-86391 DLS-SW-TRM277-F-050719 Dup E05-0210-86392 DLS-SW-TRM277-F-050719 LS E05-0210-86393 DLS-SW-TRM277-F-050719 HS 0.0111 0.0125 0.0633 0.464 (2) Field Matrix Spike % Recovery Average Result Sample/Sample Duplicate (ng/mL) %RPD Extraction Date NA 07/26/2005 NA 07/26/2005 < 0.00221 (3)n a 111 07/26/2005 115 07/26/2005 NA 07/26/2005 NA 07/26/2005 < 0.00221 (3)n a 105 07/26/2005 121 07/26/2005 NA 07/26/2005 NA 07/26/2005 < 0.00221 (3)n a 109 07/26/2005 119 07/26/2005 121 07/26/2005 108 07/26/2005 0.0147 4.78 NA 07/26/2005 NA 07/26/2005 NA 07/26/2005 NA 07/26/2005 0.0219 5.48 110 07/26/2005 97.8 07/26/2005 NA 07/26/2005 NA 07/26/2005 0.0199 2.01 112 07/26/2005 114 07/26/2005 NA 07/26/2005 NA 07/26/2005 0.0192 11.4 110 07/26/2005 111 07/26/2005 NA 07/26/2005 NA 07/26/2005 0.0119 6.72 111 07/26/2005 114 07/26/2005 NA 07/26/2005 NA 07/26/2005 0.0118 11.9 117 07/26/2005 102 07/26/2005 E05-0210 Interim R eport#13 Tennessee R iver Page 28 of 160 3M Environm ental Labora tory E05-0210 Interim R eport#13 Table 16. PFBS Sample Results Continued. 3M Sample Number Sample Description (1)PFBS Conc. (ng/mL) Average Result (2)Field Matrix Sample/Sample Spike % Duplicate Recovery (ng/mL) %RPD Extraction Date E05-0210-86394 DLS-SW-TRM271-0-050719 E05-0210-86395 DLS-SW-TRM271-0-050719 Dup E05-0210-86396 DLS-SW-TRM271-0-050719 LS E05-0210-86397 DLS-SW-TRM271-0-050719 HS 0.0133 0.0142 0.0586 0.483 NA NA 102 106 0.0138 07/26/2005 07/26/2005 6.54 07/26/2005 07/26/2005 E05-0210-86398 DLS-SW-TRM265-0-050719 E05-0210-86399 DLS-SW-TRM265-0-050719 Dup E05-0210-86400 DLS-SW-TRM265-0-050719 LS E05-0210-86401 DLS-SW-TRM265-0-050719 HS 0.0103 0.0104 0.0544 0.484 NA NA 99.9 107 0.0104 0.966 07/26/2005 07/26/2005 07/26/2005 07/26/2005 E05-0210-86402 DLS-SW-TRM261-2-050720 E05-0210-86403 DLS-SW-TRM261-2-050720 Dup E05-0210-86404 DLS-SW-TRM261-2-050720 LS E05-0210-86405 DLS-SW-TRM261-2-050720 HS < 0.00221 < 0.00221 0.0497 0.485 NA NA 113 110 < 0.00221 07/26/2005 07/26/2005 NA 07/26/2005 07/26/2005 E05-0210-86406 DLS-SW-TRM261-0-050720 E05-0210-86407 DLS-SW-TRM261-0-050720 Dup E05-0210-86408 DLS-SW-TRM261-0-050720 LS E05-0210-86409 DLS-SW-TRM261-0-050720 HS 0.0118 0.0110 0.0556 0.456 NA NA 100 101 0.0114 07/26/2005 07/26/2005 7.02 07/26/2005 07/26/2005 E05-0210-86410 DLS-SW-TRM261-F-050720 E05-0210-86411 DLS-SW-TRM261-F-050720 Dup E05-0210-86412 DLS-SW-TRM261-F-050720 LS E05-0210-86413 DLS-SW-TRM261-F-050720 HS 0.0116 0.0107 0.0621 0.460 NA NA 130 103 0.0112 07/26/2005 07/26/2005 8.07 07/26/2005 07/26/2005 E05-0210-86414 DLS-SW-TRM256-0-050720 E05-0210-86415 DLS-SW-TRM256-0-050720 Dup E05-0210-86416 DLS-SW-TRM256-0-050720 LS E05-0210-86417 DLS-SW-TRM256-0-050720 HS 0.0111 0.0125 0.0607 0.498 NA NA 111 110 0.0118 07/26/2005 07/26/2005 11.9 07/26/2005 07/26/2005 E05-0210-86418 DLS-SW-TRM254-0-050720 E05-0210-86419 DLS-SW-TRM254-0-050720 Dup E05-0210-86420 DLS-SW-TRM254-0-050720 LS E05-0210-86421 DLS-SW-TRM254-0-050720 HS 0.0124 0.0120 0.0648 0.493 NA NA 119 109 0.0122 07/26/2005 07/26/2005 3.28 07/27/2005 07/27/2005 E05-0210-86422 DLS-SW-TRM254-F-050720 E05-0210-86423 DLS-SW-TRM254-F-050720 Dup E05-0210-86424 DLS-SW-TRM254-F-050720 LS E05-0210-86425 DLS-SW-TRM254-F-050720 HS 0.0128 0.0121 0.0598 0.530 NA NA 107 112 0.0125 07/27/2005 07/27/2005 5.62 07/27/2005 07/27/2005 E05-0210-86426 DLS-SW-TRM254-2-050720 E05-0210-86427 DLS-SW-TRM254-2-050720 Dup E05-0210-86428 DLS-SW-TRM254-2-050720 LS E05-0210-86429 DLS-SW-TRM254-2-050720 HS < 0.00221 < 0.00221 0.0502 0.485 NA NA 114 110 < 0.00221 (3)n a 07/27/2005 07/27/2005 07/27/2005 07/27/2005 E05-0210-86430 DXS-SW-T01001-0-050721 E05-0210-86431 DXS-SW-T01001-0-050721 Dup E05-0210-86432 DXS-SW-T01001-0-050721 LS E05-0210-86433 DXS-SW-T01001-0-050721 HS 0.0215 0.0201 0.0707 0.519 NA NA 113 113 0.0208 07/27/2005 07/27/2005 6.73 07/27/2005 07/27/2005 E05-0210 Interim R eport#13 Tennessee R iver Page 29 of 160 3M Environm ental Labora tory E05-0210 Interim R eport#13 Table 16. PFBS Sample Results Continued. 3M Sample Number Sample Description E05-0210-86434 DXS-SW-T01002-0-050721 E05-0210-86435 DXS-SW-T01002-0-050721 Dup E05-0210-86436 DXS-SW-T01002-0-050721 LS E05-0210-86437 DXS-SW-T01002-0-050721 HS E05-0210-86438 DXS-SW-T01003-0-050721 E05-0210-86439 DXS-SW-T01003-0-050721 Dup E05-0210-86440 DXS-SW-T01003-0-050721 LS E05-0210-86441 DXS-SW-T01003-0-050721 HS E05-0210-86442 DXS-SW-T01004-0-050721 E05-0210-86443 DXS-SW-T01004-0-050721 Dup E05-0210-86444 DXS-SW-T01004-0-050721 LS E05-0210-86445 DXS-SW-T01004-0-050721 HS E05-0210-86446 DXS-SW-T01005-0-050721 E05-0210-86447 DXS-SW-T01005-0-050721 Dup E05-0210-86448 DXS-SW-T01005-0-050721 LS E05-0210-86449 DXS-SW-T01005-0-050721 HS E05-0210-86450 DXS-SW-T01006-0-050721 E05-0210-86451 DXS-SW-T01006-0-050721 Dup E05-0210-86452 DXS-SW-T01006-0-050721 LS E05-0210-86453 DXS-SW-T01006-0-050721 HS E05-0210-86454 DXS-SW-T01007-0-050721 E05-0210-86455 DXS-SW-T01007-0-050721 Dup E05-0210-86456 DXS-SW-T01007-0-050721 LS E05-0210-86457 DXS-SW-T01007-0-050721 HS E05-0210-86458 DXS-SW-T01008-0-050721 E05-0210-86459 DXS-SW-T01008-0-050721 Dup E05-0210-86460 DXS-SW-T01008-0-050721 LS E05-0210-86461 DXS-SW-T01008-0-050721 HS E05-0210-86462 DXS-SW-T01009-0-050721 E05-0210-86463 DXS-SW-T01009-0-050721 Dup E05-0210-86464 DXS-SW-T01009-0-050721 LS E05-0210-86465 DXS-SW-T01009-0-050721 HS E05-0210-86466 DXS-SW-T01010-0-050721 E05-0210-86467 DXS-SW-T01010-0-050721 Dup E05-0210-86468 DXS-SW-T01010-0-050721 LS E05-0210-86469 DXS-SW-T01010-0-050721 HS E05-0210-86470 DXS-SW-T02001-0-050721 E05-0210-86471 DXS-SW-T02001-0-050721 Dup E05-0210-86472 DXS-SW-T02001-0-050721 LS E05-0210-86473 DXS-SW-T02001-0-050721 HS (1)PFBS Conc. (ng/mL) 0.0161 0.0164 0.0663 0.521 0.0165 0.0166 0.0624 0.460 0.0158 0.0165 0.0678 0.470 0.00957 0.00985 0.0863 0.482 0.00432 0.00366 0.0646 0.546 0.00566 0.00564 0.1040 0.485 0.00369 0.00502 0.0635 0.452 0.00491 0.00444 0.0532 0.535 0.00395 0.00385 0.0543 0.501 0.0414 0.0423 0.0856 0.499 Average Result (2)Field Matrix Sample/Sample Spike % Duplicate Recovery (ng/mL) %RPD Extraction Date NA 07/27/2005 NA 07/27/2005 0.0163 1.85 113 07/27/2005 114 07/27/2005 NA 07/27/2005 NA 07/27/2005 104 0.0166 0.604 07/27/2005 104 07/28/2005 c e n trifu g e d NA 07/27/2005 NA 07/27/2005 0.0162 4.33 117 07/27/2005 103 07/27/2005 NA 07/27/2005 NA 07/27/2005 0.00971 2.88 174 07/27/2005 107 07/27/2005 NA 07/27/2005 NA 07/27/2005 0.00399 16.5 137 07/27/2005 123 07/27/2005 NA 07/27/2005 NA 07/27/2005 0.00565 0.354 223 07/27/2005 109 07/27/2005 NA 07/27/2005 NA 07/27/2005 0.00436 30.5 134 07/27/2005 102 07/27/2005 NA 07/27/2005 NA 07/27/2005 0.00468 10.0 110 07/27/2005 120 07/27/2005 NA 07/27/2005 NA 07/27/2005 0.00390 2.56 114 07/27/2005 113 07/27/2005 NA NA 99.2 104 0.0419 07/27/2005 07/27/2005 2.15 07/27/2005 07/27/2005 E05-0210 Interim R eport#13 Tennessee R iver Page 30 of 160 3M Environm ental Labora tory E05-0210 Interim R eport#13 Table 16. PFBS Sample Results Continued. 3M Sample Number Sample Description (1)PFBS Conc. (ng/mL) Average (2>Field Matrix Result Spike % Sample/Sample Recovery Duplicate %RPD Extraction Date E05-0210-86474 DXS-SW-T02002-0-050721 E05-0210-86475 DXS-SW-T02002-0-050721 Dup E05-0210-86476 DXS-SW-T02002-0-050721 LS E05-0210-86477 DXS-SW-T02002-0-050721 HS 0.0306 0.0316 0.0783 0.508 NA NA 107 108 0.0311 07/27/2005 07/27/2005 3.22 07/27/2005 07/27/2005 E05-0210-86478 DXS-SW-T02003-0-050721 E05-0210-86479 DXS-SW-T02003-0-050721 Dup E05-0210-86480 DXS-SW-T02003-0-050721 LS E05-0210-86481 DXS-SW-T02003-0-050721 HS < 0.00221 < 0.00221 0.0479 0.476 NA NA 109 108 < 0.00221 (3)n a 07/27/2005 07/27/2005 07/27/2005 07/27/2005 E05-0210-86482 DXS-SW-T02004-0-050721 E05-0210-86483 DXS-SW-T02004-0-050721 Dup E05-0210-86484 DXS-SW-T02004-0-050721 LS E05-0210-86485 DXS-SW-T02004-0-050721 HS < 0.00221 < 0.00221 0.0544 0.476 NA NA 122 108 < 0.00221 (3)n a 07/27/2005 07/27/2005 07/27/2005 07/27/2005 E05-0210-86486 DXS-SW-T02005-0-050721 E05-0210-86487 DXS-SW-T02005-0-050721 Dup E05-0210-86488 DXS-SW-T02005-0-050721 LS E05-0210-86489 DXS-SW-T02005-0-050721 HS < 0.000883 < 0.00221 0.0511 0.600 NA NA 116 136 < 0.00221 (3)n a 07/27/2005 07/28/2005 c e n trifu g e d 07/27/2005 07/28/2005 c e n trifu g e d E05-0210-86490 DXS-SW-T02006-0-050721 E05-0210-86491 DXS-SW-T02006-0-050721 Dup E05-0210-86492 DXS-SW-T02006-0-050721 LS E05-0210-86493 DXS-SW-T02006-0-050721 HS < 0.000883 0.00101 0.0534 0.545 NA NA 119 123 (5)0.00101 07/27/2005 07/27/2005 NA 07/27/2005 07/27/2005 E05-0210-86494 DXS-SW-T02007-0-050721 E05-0210-86495 DXS-SW-T02007-0-050721 Dup E05-0210-86496 DXS-SW-T02007-0-050721 LS E05-0210-86497 DXS-SW-T02007-0-050721 HS < 0.000883 < 0.000883 0.0470 0.450 NA NA 106 102 < 0.000883 (3)n a 07/27/2005 07/27/2005 07/27/2005 07/27/2005 E05-0210-86498 DXS-SW-T02008-0-050721 E05-0210-86499 DXS-SW-T02008-0-050721 Dup E05-0210-86500 DXS-SW-T02008-0-050721 LS E05-0210-86501 DXS-SW-T02008-0-050721 HS 0.00109 0.00115 0.0508 0.468 NA NA 112 106 0.00112 07/27/2005 07/27/2005 5.36 07/27/2005 07/27/2005 E05-0210-86502 DXS-SW-T02009-0-050721 E05-0210-86503 DXS-SW-T02009-0-050721 Dup E05-0210-86504 DXS-SW-T02009-0-050721 LS E05-0210-86505 DXS-SW-T02009-0-050721 HS 0.00288 0.00244 0.0541 0.498 NA NA 117 112 0.00266 07/27/2005 07/27/2005 16.5 07/27/2005 07/27/2005 E05-0210-86506 DXS-SW-T02010-0-050721 E05-0210-86507 DXS-SW-T02010-0-050721 Dup E05-0210-86508 DXS-SW-T02010-0-050721 LS E05-0210-86509 DXS-SW-T02010-0-050721 HS 0.00628 0.00553 0.0533 0.344 NA NA 107 76.7 0.00591 07/27/2005 07/27/2005 12.7 07/27/2005 07/27/2005 E05-0210-86510 DXS-SW-T03001-0-050721 E05-0210-86511 DXS-SW-T03001-0-050721 Dup E05-0210-86512 DXS-SW-T03001-0-050721 LS E05-0210-86513 DXS-SW-T03001-0-050721 HS 0.818 0.854 0.911 1.33 NA NA (6)n r 112 0.836 07/27/2005 4.31 07/27/2005 07/27/2005 07/27/2005 E05-0210 Interim R eport#13 Tennessee R iver Page 31 of 160 3M Environm ental Laboratory E05-0210 Interim R eport#13 Table 16. PFBS Sample Results Continued. 3M Sample Number Sample Description (1)PFBS Conc. (ng/mL) Average (2)Field Matrix Result Spike % Sample/Sample Recovery Duplicate %RPD Extraction Date E05-0210-86514 DXS-SW-T03002-0-050721 E05-0210-86515 DXS-SW-T03002-0-050721 Dup E05-0210-86516 DXS-SW-T03002-0-050721 LS E05-0210-86517 DXS-SW-T03002-0-050721 HS 0.00519 0.00424 0.0521 0.480 NA NA 107 108 0.00472 07/27/2005 07/27/2005 20.1 07/27/2005 07/27/2005 E05-0210-86518 DXS-SW-T03003-0-050721 E05-0210-86519 DXS-SW-T03003-0-050721 Dup E05-0210-86520 DXS-SW-T03003-0-050721 LS E05-0210-86521 DXS-SW-T03003-0-050721 HS < 0.000883 < 0.000883 0.0519 0.487 NA NA 118 110 < 0.000883 (3)n a 07/27/2005 07/27/2005 07/27/2005 07/27/2005 E05-0210-86522 DXS-SW-T03004-0-050722 E05-0210-86523 DXS-SW-T03004-0-050722 Dup E05-0210-86524 DXS-SW-T03004-0-050722 LS E05-0210-86525 DXS-SW-T03004-0-050722 HS 0.00430 0.00106 0.0536 0.516 NA NA 115 116 0.00268 07/27/2005 07/27/2005 121 07/27/2005 07/27/2005 E05-0210-86526 DXS-SW-T03005-0-050722 E05-0210-86527 DXS-SW-T03005-0-050722 Dup E05-0210-86528 DXS-SW-T03005-0-050722 LS E05-0210-86529 DXS-SW-T03005-0-050722 HS < 0.000883 0.00171 0.0588 0.536 NA NA 133 122 (5)0.00171 07/27/2005 07/28/2005 NA c e n trifu g e d 07/27/2005 07/27/2005 E05-0210-86530 DXS-SW-T03006-0-050722 E05-0210-86531 DXS-SW-T03006-0-050722 Dup E05-0210-86532 DXS-SW-T03006-0-050722 LS E05-0210-86533 DXS-SW-T03006-0-050722 HS < 0.000883 < 0.000883 0.0430 0.591 NA NA 97.5 134 < 0.000883 (3)n a 07/27/2005 07/27/2005 07/28/2005 dc e n t r i f u g e 07/27/2005 E05-0210-86534 DXS-SW-T03007-0-050722 E05-0210-86535 DXS-SW-T03007-0-050722 Dup E05-0210-86536 DXS-SW-T03007-0-050722 LS E05-0210-86537 DXS-SW-T03007-0-050722 HS < 0.000883 0.00107 0.0564 0.513 NA NA 128 116 (5)0.00107 07/27/2005 07/28/2005 NA c e n trifu g e d 07/27/2005 07/27/2005 E05-0210-86538 DXS-SW-T03008-0-050722 E05-0210-86539 DXS-SW-T03008-0-050722 Dup E05-0210-86540 DXS-SW-T03008-0-050722 LS E05-0210-86541 DXS-SW-T03008-0-050722 HS < 0.000883 < 0.000883 0.0513 0.507 NA NA 116 115 < 0.000883 (3)n a 07/27/2005 07/27/2005 07/27/2005 07/27/2005 E05-0210-86542 DXS-SW-T03009-0-050722 E05-0210-86543 DXS-SW-T03009-0-050722 Dup E05-0210-86544 DXS-SW-T03009-0-050722 LS E05-0210-86545 DXS-SW-T03009-0-050722 HS < 0.000883 < 0.000883 0.0505 0.460 NA NA 114 104 < 0.000883 (3)n a 07/27/2005 07/27/2005 07/27/2005 07/27/2005 E05-0210-86546 DXS-SW-T03010-0-050722 E05-0210-86547 DXS-SW-T03010-0-050722 Dup E05-0210-86548 DXS-SW-T03010-0-050722 LS E05-0210-86549 DXS-SW-T03010-0-050722 HS 0.000908 < 0.000883 0.0498 0.449 NA NA 111 102 (5)0.000908 07/27/2005 07/27/2005 NA 07/27/2005 07/27/2005 E05-0210 Interim R eport#13 Tennessee R iver Page 32 of 160 3M Environm ental Laboratory E05-0210 Interim R eport#13 Table 16. PFBS Sample Results Continued. 3 M Sam ple N um ber Sam ple Description (1)P F B S Conc. (ng/m L) A v e ra g e (2)F ie ld M a tr ix R e s u lt Spike % Sam ple/Sam ple Recovery D u p lic a te % RPD Extraction Date E05-0210-86550 DLS-SW-TRIP01 E05-0210-86551 DLS-SW-TRIP01 LS E05-0210-86552 DLS-SW-TRIP01 HS < 0.000883 0.04710 0.444 NA 107 101 < 0.000883 07/27/2005 NA 07/27/2005 07/27/2005 E05-0210-86553 DXS-SW-TRIP02 E05-0210-86554 DXS-SW-TRIP02 LS E05-0210-86555 DXS-SW-TRIP02 HS < 0.000883 0.0478 0.440 NA 108 99.8 < 0.000883 07/27/2005 NA 07/27/2005 07/27/2005 E05-0210-86556 DXS-SW-TRIP03 E05-0210-86557 DXS-SW-TRIP03 LS E05-0210-86558 DXS-SW-TRIP03 HS < 0.000883 0.0495 0.080 NA 112 18.0 < 0.000883 07/27/2005 NA 07/27/2005 07/27/2005 (1) The analytical uncertainty of the PFBS resuts is 10010.6% based on method accuracy and precision. All results and calculations in this table are presented with three significant figures. Sample concentrations, averages, and %RPD values may vary slightly from the raw data. (2) Low field matrix spike concentration = 0.0441 ng/mL. High field matrix spike concentration = 0.441 ng/mL. (3) NA: Not applicable. %RPD value was not determined when concentrations for both the sample and sample duplicate were below the stated LOQ. (4) Sample descriptions were misidentified when checked into LIMS. Samples with LIMS ID numbers 86370 and 86371 were in bottles labeled low spike and high spike, respectively. Bottles with LIMS ID numbers 86372 and 86372 were surrogate spiked only (sample and sample duplicate bottles). (5) The average value listed is the concentration for the sample or sample duplicate that produced a value above the LOQ. A true average and %RPD between the sample and sample duplicate was not determined. (6) NR: Not reported. Spike recovery not calculated as the endogenous sample concentration was at least three times the spiked amount. E05-0210 Interim R eport#13 Tennessee R iver Page 33 of 160 3M Environm ental Labora tory E05-0210 Interim R eport#13 Table 17. PFHS Sample Results. 3M Sample Number Sample Description (1)PFHS Conc. (ng/mL) Average (2>Field Matrix Result Spike % Sample/Sample Recovery Duplicate %RPD Extraction Date E05-0210-86358 DLS-SW-TRM307-0-050718 < 0.00104 NA 07/26/2005 E05-0210-86359 DLS-SW-TRM307-0-050718 Dup E05-0210-86360 DLS-SW-TRM307-0-050718 LS < 0.00104 0.0581 NA 112 < 0.00104 (3)n a 07/26/2005 07/26/2005 E05-0210-86361 DLS-SW-TRM307-0-050718 HS 0.584 112 07/26/2005 E05-0210-86362 DLS-SW-TRM301-0-050718 < 0.00104 NA 07/26/2005 E05-0210-86363 DLS-SW-TRM301-0-050718 Dup E05-0210-86364 DLS-SW-TRM301-0-050718 LS 0.00134 0.0542 NA 101 0.00134 (4)n a 07/26/2005 07/26/2005 E05-0210-86365 DLS-SW-TRM301-0-050718 HS 0.600 115 07/26/2005 E05-0210-86366 DLS-SW-TRM301-F-050718 < 0.00104 NA 07/26/2005 E05-0210-86367 DLS-SW-TRM301-F-050718 Dup E05-0210-86368 DLS-SW-TRM301-F-050718 LS < 0.00104 0.0577 NA 111 < 0.00104 (3)n a 07/26/2005 07/26/2005 E05-0210-86369 DLS-SW-TRM301-F-050718 HS 0.576 110 07/26/2005 E05-0210-86370 DLS-SW-TRM295-0-050719(5) 0.0719 116 07/26/2005 E05-0210-86371 DLS-SW-TRM295-0-050719 Dup(5) E05-0210-86372 DLS-SW-TRM295-0-050719 LS(5) 0.5490 0.0115 103 NA 0.0117 07/26/2005 2.58 07/26/2005 E05-0210-86373 DLS-SW-TRM295-0-050719 HS(5) 0.0118 NA 07/26/2005 E05-0210-86374 DLS-SW-TRM289-0-050719 0.0133 NA 07/26/2005 E05-0210-86375 DLS-SW-TRM289-0-050719 Dup E05-0210-86376 DLS-SW-TRM289-0-050719 LS 0.0123 0.0666 NA 103 0.01280 07/26/2005 7.81 07/26/2005 E05-0210-86377 DLS-SW-TRM289-0-050719 HS 0.493 92.2 07/26/2005 E05-0210-86378 DLS-SW-TRM289-F-050719 0.0129 NA 07/26/2005 E05-0210-86379 DLS-SW-TRM289-F-050719 Dup E05-0210-86380 DLS-SW-TRM289-F-050719 LS 0.0120 0.0706 NA 112 0.01245 07/26/2005 7.23 07/26/2005 E05-0210-86381 DLS-SW-TRM289-F-050719 HS 0.582 109 07/26/2005 E05-0210-86382 DLS-SW-TRM283-0-050719 0.00888 NA 07/26/2005 E05-0210-86383 DLS-SW-TRM283-0-050719 Dup E05-0210-86384 DLS-SW-TRM283-0-050719 LS 0.00883 0.0645 NA 107 0.00886 0.565 07/26/2005 07/26/2005 E05-0210-86385 DLS-SW-TRM283-0-050719 HS 0.572 108 07/26/2005 E05-0210-86386 DLS-SW-TRM277-0-050719 0.00579 NA 07/26/2005 E05-0210-86387 DLS-SW-TRM277-0-050719 Dup E05-0210-86388 DLS-SW-TRM277-0-050719 LS 0.00704 0.0637 NA 110 0.00642 07/26/2005 19.5 07/26/2005 E05-0210-86389 DLS-SW-TRM277-0-050719 HS 0.574 109 07/26/2005 E05-0210 Interim R eport#13 Tennessee R iver Page 34 of 160 3M Environm ental Labora tory E05-0210 Interim R eport#13 Table 17. PFHS Sample Results Continued. 3M Sample Number Sample Description (1)PFHS Conc. (ng/mL) Average (2>Field Matrix Result Spike % Sample/Sample Recovery Duplicate %RPD Extraction Date E05-0210-86390 DLS-SW-TRM277-F-050719 0.00609 NA 07/26/2005 E05-0210-86391 DLS-SW-TRM277-F-050719 Dup E05-0210-86392 DLS-SW-TRM277-F-050719 LS 0.00672 0.0637 NA 110 0.00641 9.84 07/26/2005 07/26/2005 E05-0210-86393 DLS-SW-TRM277-F-050719 HS 0.529 100 07/26/2005 E05-0210-86394 DLS-SW-TRM271-0-050719 0.00512 NA 07/26/2005 E05-0210-86395 DLS-SW-TRM271-0-050719 Dup E05-0210-86396 DLS-SW-TRM271-0-050719 LS 0.00574 0.0573 NA 99.6 0.00543 07/26/2005 11.4 07/26/2005 E05-0210-86397 DLS-SW-TRM271-0-050719 HS 0.558 106 07/26/2005 E05-0210-86398 DLS-SW-TRM265-0-050719 0.00525 NA 07/26/2005 E05-0210-86399 DLS-SW-TRM265-0-050719 Dup E05-0210-86400 DLS-SW-TRM265-0-050719 LS 0.00542 0.0568 NA 98.8 0.00534 07/26/2005 3.19 07/26/2005 E05-0210-86401 DLS-SW-TRM265-0-050719 HS 0.561 107 07/26/2005 E05-0210-86402 DLS-SW-TRM261-2-050720 < 0.00104 NA 07/26/2005 E05-0210-86403 DLS-SW-TRM261-2-050720 Dup E05-0210-86404 DLS-SW-TRM261-2-050720 LS < 0.00104 0.0604 NA 116 < 0.00104 (3)n a 07/26/2005 07/26/2005 E05-0210-86405 DLS-SW-TRM261-2-050720 HS 0.576 110 07/26/2005 E05-0210-86406 DLS-SW-TRM261-0-050720 0.00520 NA 07/26/2005 E05-0210-86407 DLS-SW-TRM261-0-050720 Dup E05-0210-86408 DLS-SW-TRM261-0-050720 LS 0.00481 0.0568 NA 99.4 0.00501 07/26/2005 7.79 07/26/2005 E05-0210-86409 DLS-SW-TRM261-0-050720 HS 0.530 101 07/26/2005 E05-0210-86410 DLS-SW-TRM261-F-050720 0.00542 NA 07/26/2005 E05-0210-86411 DLS-SW-TRM261-F-050720 Dup E05-0210-86412 DLS-SW-TRM261-F-050720 LS 0.00486 0.0586 NA 103 0.00514 07/26/2005 10.9 07/27/2005 E05-0210-86413 DLS-SW-TRM261-F-050720 HS 0.512 97.3 07/27/2005 E05-0210-86414 DLS-SW-TRM256-0-050720 0.00485 NA 07/27/2005 E05-0210-86415 DLS-SW-TRM256-0-050720 Dup E05-0210-86416 DLS-SW-TRM256-0-050720 LS 0.00470 0.0611 NA 108 0.00478 3.14 07/27/2005 07/27/2005 E05-0210-86417 DLS-SW-TRM256-0-050720 HS 0.559 106 07/27/2005 E05-0210-86418 DLS-SW-TRM254-0-050720 0.00550 NA 07/27/2005 E05-0210-86419 DLS-SW-TRM254-0-050720 Dup E05-0210-86420 DLS-SW-TRM254-0-050720 LS 0.00481 0.0623 NA 110 0.00516 07/27/2005 13.4 07/27/2005 E05-0210-86421 DLS-SW-TRM254-0-050720 HS 0.555 106 07/27/2005 E05-0210-86422 DLS-SW-TRM254-F-050720 0.00533 NA 07/27/2005 E05-0210-86423 DLS-SW-TRM254-F-050720 Dup E05-0210-86424 DLS-SW-TRM254-F-050720 LS 0.00525 0.0639 NA 112 0.00529 07/27/2005 1.51 07/27/2005 E05-0210-86425 DLS-SW-TRM254-F-050720 HS 0.585 111 07/27/2005 E05-0210 Interim R eport#13 Tennessee R iver Page 35 of 160 3M Environm ental Labora tory E05-0210 Interim R eport#13 Table 17. PFHS Sample Results Continued 3M Sample Number Sample Description (1)PFHS Conc. (ng/mL) Average (2>Field Matrix Result Spike % Sample/Sample Recovery Duplicate %RPD Extraction Date E05-0210-86426 DLS-SW-TRM254-2-050720 < 0.00104 NA 07/27/2005 E05-0210-86427 DLS-SW-TRM254-2-050720 Dup E05-0210-86428 DLS-SW-TRM254-2-050720 LS < 0.00104 0.0578 NA 111 < 0.00104 (3)n a 07/27/2005 07/27/2005 E05-0210-86429 DLS-SW-TRM254-2-050720 HS 0.575 110 07/27/2005 E05-0210-86430 DXS-SW-T01001-0-050721 0.0121 NA 07/27/2005 E05-0210-86431 DXS-SW-T01001-0-050721 Dup E05-0210-86432 DXS-SW-T01001-0-050721 LS 0.0116 0.0691 NA 110 0.0119 4.22 07/27/2005 07/27/2005 E05-0210-86433 DXS-SW-T01001-0-050721 HS 0.570 107 07/27/2005 E05-0210-86434 DXS-SW-T01002-0-050721 0.0101 NA 07/27/2005 E05-0210-86435 DXS-SW-T01002-0-050721 Dup E05-0210-86436 DXS-SW-T01002-0-050721 LS 0.00959 0.0637 NA 103 0.00985 07/27/2005 5.18 07/27/2005 E05-0210-86437 DXS-SW-T01002-0-050721 HS 0.593 112 07/27/2005 E05-0210-86438 DXS-SW-T01003-0-050721 0.00827 NA 07/27/2005 E05-0210-86439 DXS-SW-T01003-0-050721 Dup E05-0210-86440 DXS-SW-T01003-0-050721 LS 0.00941 0.0625 NA 103 0.00884 07/27/2005 12.9 07/27/2005 E05-0210-86441 DXS-SW-T01003-0-050721 HS 0.517 99.2 07/28/2005 centrifuged E05-0210-86442 DXS-SW-T01004-0-050721 0.0097 NA 07/27/2005 E05-0210-86443 DXS-SW-T01004-0-050721 Dup E05-0210-86444 DXS-SW-T01004-0-050721 LS 0.0101 0.0692 NA 114 0.00989 4.35 07/27/2005 07/27/2005 E05-0210-86445 DXS-SW-T01004-0-050721 HS 0.512 96.4 07/27/2005 E05-0210-86446 DXS-SW-T01005-0-050721 0.00633 NA 07/27/2005 E05-0210-86447 DXS-SW-T01005-0-050721 Dup E05-0210-86448 DXS-SW-T01005-0-050721 LS 0.00667 0.0951 NA 170 0.00650 07/27/2005 5.23 07/27/2005 E05-0210-86449 DXS-SW-T01005-0-050721 HS 0.541 102 07/27/2005 E05-0210-86450 DXS-SW-T01006-0-050721 0.00346 NA 07/27/2005 E05-0210-86451 DXS-SW-T01006-0-050721 Dup E05-0210-86452 DXS-SW-T01006-0-050721 LS 0.00333 0.0731 NA 134 0.00340 07/27/2005 3.83 07/27/2005 E05-0210-86453 DXS-SW-T01006-0-050721 HS 0.611 117 07/27/2005 E05-0210-86454 DXS-SW-T01007-0-050721 0.00419 NA 07/27/2005 E05-0210-86455 DXS-SW-T01007-0-050721 Dup E05-0210-86456 DXS-SW-T01007-0-050721 LS 0.00429 0.0628 NA 112 0.00424 07/27/2005 2.36 07/27/2005 E05-0210-86457 DXS-SW-T01007-0-050721 HS 0.559 106 07/27/2005 E05-0210-86458 DXS-SW-T01008-0-050721 0.00316 NA 07/27/2005 E05-0210-86459 DXS-SW-T01008-0-050721 Dup E05-0210-86460 DXS-SW-T01008-0-050721 LS 0.00328 0.0686 NA 125 0.00322 07/27/2005 3.73 07/27/2005 E05-0210-86461 DXS-SW-T01008-0-050721 HS 0.507 96.7 07/27/2005 E05-0210 Interim R eport#13 Tennessee R iver Page 36 of 160 3M Environm ental Laboratory E05-0210 Interim R eport#13 Table 17. PFHS Sample Results Continued. 3M Sample Number Sample Description (1)PFHS Conc. (ng/mL) Average (2)Field Matrix Result Spike % Sample/Sample Recovery Duplicate %RPD Extraction Date E05-0210-86462 DXS-SW-T01009-0-050721 0.00301 NA 07/27/2005 E05-0210-86463 DXS-SW-T01009-0-050721 Dup E05-0210-86464 DXS-SW-T01009-0-050721 LS 0.00330 0.0595 NA 108 0.00316 07/27/2005 9.19 07/27/2005 E05-0210-86465 DXS-SW-T01009-0-050721 HS 0.601 115 07/27/2005 E05-0210-86466 DXS-SW-T01010-0-050721 0.00332 NA 07/27/2005 E05-0210-86467 DXS-SW-T01010-0-050721 Dup E05-0210-86468 DXS-SW-T01010-0-050721 LS 0.00277 0.0607 NA 111 0.00305 07/27/2005 18.1 07/27/2005 E05-0210-86469 DXS-SW-T01010-0-050721 HS 0.561 107 07/27/2005 E05-0210-86470 DXS-SW-T02001-0-050721 0.0205 NA 07/27/2005 E05-0210-86471 DXS-SW-T02001-0-050721 Dup E05-0210-86472 DXS-SW-T02001-0-050721 LS 0.0187 0.0729 NA 102 0.0196 07/27/2005 9.18 07/27/2005 E05-0210-86473 DXS-SW-T02001-0-050721 HS 0.560 104 07/27/2005 E05-0210-86474 DXS-SW-T02002-0-050721 0.0124 NA 07/27/2005 E05-0210-86475 DXS-SW-T02002-0-050721 Dup E05-0210-86476 DXS-SW-T02002-0-050721 LS 0.0126 0.0661 NA 103 0.0125 07/27/2005 1.60 07/27/2005 E05-0210-86477 DXS-SW-T02002-0-050721 HS 0.552 104 07/27/2005 E05-0210-86478 DXS-SW-T02003-0-050721 < 0.00104 NA 07/27/2005 E05-0210-86479 DXS-SW-T02003-0-050721 Dup E05-0210-86480 DXS-SW-T02003-0-050721 LS < 0.00104 0.0553 NA 106 < 0.00104 (3)n a 07/27/2005 07/27/2005 E05-0210-86481 DXS-SW-T02003-0-050721 HS 0.528 101 07/27/2005 E05-0210-86482 DXS-SW-T02004-0-050721 < 0.00104 NA 07/27/2005 E05-0210-86483 DXS-SW-T02004-0-050721 Dup E05-0210-86484 DXS-SW-T02004-0-050721 LS < 0.00104 0.0619 NA 117 < 0.00104 (3)n a 07/27/2005 07/27/2005 E05-0210-86485 DXS-SW-T02004-0-050721 HS 0.534 102 07/27/2005 E05-0210-86486 DXS-SW-T02005-0-050721 < 0.0026 NA 07/27/2005 E05-0210-86487 DXS-SW-T02005-0-050721 Dup E05-0210-86488 DXS-SW-T02005-0-050721 LS < 0.0026 0.0544 NA 104 < 0.0026 (3)n a 07/28/2005 centrifuged 07/27/2005 E05-0210-86489 DXS-SW-T02005-0-050721 HS 0.675 119 07/28/2005 centrifuged E05-0210-86490 DXS-SW-T02006-0-050721 < 0.0026 NA 07/27/2005 E05-0210-86491 DXS-SW-T02006-0-050721 Dup E05-0210-86492 DXS-SW-T02006-0-050721 LS < 0.0026 0.0595 NA 114 < 0.0026 (3)n a 07/27/2005 07/27/2005 E05-0210-86493 DXS-SW-T02006-0-050721 HS 0.617 118 07/27/2005 E05-0210-86494 DXS-SW-T02007-0-050721 < 0.0026 NA 07/27/2005 E05-0210-86495 DXS-SW-T02007-0-050721 Dup < 0.0026 NA < 0.0026 (3)n a 07/27/2005 E05-0210-86496 DXS-SW-T02007-0-050721 LS E05-0210-86497 DXS-SW-T02007-0-050721 HS 0.0529 0.526 102 101 07/27/2005 07/27/2005 E05-0210 Interim R eport#13 Tennessee R iver Page 37 of 160 3M Environm ental Labora tory E05-0210 Interim R eport#13 Table 17. PFHS Sample Results Continued. 3M Sample Number Sample Description (1)PFHS Conc. (ng/mL) Average (2>Field Matrix Result Spike % Sample/Sample Recovery Duplicate %RPD Extraction Date E05-0210-86498 DXS-SW-T02008-0-050721 < 0.0026 NA 07/27/2005 E05-0210-86499 DXS-SW-T02008-0-050721 Dup E05-0210-86500 DXS-SW-T02008-0-050721 LS < 0.0026 0.0578 NA 111 < 0.0026 (3)n a 07/27/2005 07/27/2005 E05-0210-86501 DXS-SW-T02008-0-050721 HS 0.528 101 07/27/2005 E05-0210-86502 DXS-SW-T02009-0-050721 < 0.0026 NA 07/27/2005 E05-0210-86503 DXS-SW-T02009-0-050721 Dup E05-0210-86504 DXS-SW-T02009-0-050721 LS < 0.0026 0.0608 NA 117 < 0.0026 (3)n a 07/27/2005 07/27/2005 E05-0210-86505 DXS-SW-T02009-0-050721 HS 0.570 109 07/27/2005 E05-0210-86506 DXS-SW-T02010-0-050721 0.00363 NA 07/27/2005 E05-0210-86507 DXS-SW-T02010-0-050721 Dup E05-0210-86508 DXS-SW-T02010-0-050721 LS 0.00342 0.0554 NA 99.6 0.00353 07/27/2005 5.96 07/27/2005 E05-0210-86509 DXS-SW-T02010-0-050721 HS 0.357 67.8 07/27/2005 E05-0210-86510 DXS-SW-T03001-0-050721 0.176 NA 07/27/2005 E05-0210-86511 DXS-SW-T03001-0-050721 Dup E05-0210-86512 DXS-SW-T03001-0-050721 LS 0.185 0.250 NA 133 0.181 4.99 07/27/2005 07/27/2005 E05-0210-86513 DXS-SW-T03001-0-050721 HS 0.742 108 07/27/2005 E05-0210-86514 DXS-SW-T03002-0-050721 < 0.0026 NA 07/27/2005 E05-0210-86515 DXS-SW-T03002-0-050721 Dup E05-0210-86516 DXS-SW-T03002-0-050721 LS < 0.0026 0.0568 NA 109 < 0.0026 (3)n a 07/27/2005 07/27/2005 E05-0210-86517 DXS-SW-T03002-0-050721 HS 0.549 105 07/27/2005 E05-0210-86518 DXS-SW-T03003-0-050721 < 0.0026 NA 07/27/2005 E05-0210-86519 DXS-SW-T03003-0-050721 Dup E05-0210-86520 DXS-SW-T03003-0-050721 LS < 0.0026 0.0581 NA 112 < 0.0026 (3)n a 07/27/2005 07/27/2005 E05-0210-86521 DXS-SW-T03003-0-050721 HS 0.558 107 07/27/2005 E05-0210-86522 DXS-SW-T03004-0-050722 0.00756 NA 07/27/2005 E05-0210-86523 DXS-SW-T03004-0-050722 Dup E05-0210-86524 DXS-SW-T03004-0-050722 LS < 0.0026 0.0597 NA 100 (4)0.00756 07/27/2005 NA 07/27/2005 E05-0210-86525 DXS-SW-T03004-0-050722 HS 0.570 108 07/27/2005 E05-0210-86526 DXS-SW-T03005-0-050722 < 0.0026 NA 07/27/2005 E05-0210-86527 DXS-SW-T03005-0-050722 Dup E05-0210-86528 DXS-SW-T03005-0-050722 LS 0.00331 0.0649 NA 118 (4)0.00331 07/28/2005 NA c e n trifu g e d 07/27/2005 E05-0210-86529 DXS-SW-T03005-0-050722 HS 0.589 112 07/27/2005 E05-0210-86530 DXS-SW-T03006-0-050722 < 0.0026 NA 07/27/2005 E05-0210-86531 DXS-SW-T03006-0-050722 Dup E05-0210-86532 DXS-SW-T03006-0-050722 LS E05-0210-86533 DXS-SW-T03006-0-050722 HS < 0.0026 0.0285 0.643 NA 54.7 123 < 0.0026 (3)n a 07/27/2005 07/28/2005 c e n trifu g e d 07/27/2005 E05-0210 Interim R eport#13 Tennessee R iver Page 38 of 160 3M Environm ental Labora tory E05-0210 Interim R eport#13 Table 17. PFHS Sample Results Continued. 3M Sample Number Sample Description (1)PFHS Conc. (ng/mL) Average (2)Field Matrix Result Spike % Sample/Sample Recovery Duplicate %RPD Extraction Date E05-0210-86534 DXS-SW-T03007-0-050722 < 0.0026 NA 07/27/2005 E05-0210-86535 DXS-SW-T03007-0-050722 Dup E05-0210-86536 DXS-SW-T03007-0-050722 LS 0.00366 0.0640 NA 123 (4)0.00366 07/28/2005 NA centrifuged 07/27/2005 E05-0210-86537 DXS-SW-T03007-0-050722 HS 0.579 111 07/27/2005 E05-0210-86538 DXS-SW-T03008-0-050722 < 0.0026 NA 07/27/2005 E05-0210-86539 DXS-SW-T03008-0-050722 Dup E05-0210-86540 DXS-SW-T03008-0-050722 LS < 0.0026 0.0551 NA 106 < 0.0026 (3)n a 07/27/2005 07/27/2005 E05-0210-86541 DXS-SW-T03008-0-050722 HS 0.567 109 07/27/2005 E05-0210-86542 DXS-SW-T03009-0-050722 < 0.0026 NA 07/27/2005 E05-0210-86543 DXS-SW-T03009-0-050722 Dup E05-0210-86544 DXS-SW-T03009-0-050722 LS < 0.0026 0.0568 NA 109 < 0.0026 (3)n a 07/27/2005 07/27/2005 E05-0210-86545 DXS-SW-T03009-0-050722 HS 0.530 102 07/27/2005 E05-0210-86546 DXS-SW-T03010-0-050722 < 0.0026 NA 07/27/2005 E05-0210-86547 DXS-SW-T03010-0-050722 Dup E05-0210-86548 DXS-SW-T03010-0-050722 LS < 0.0026 0.0547 NA 105 < 0.0026 (3)NA 07/27/2005 07/27/2005 E05-0210-86549 DXS-SW-T03010-0-050722 HS 0.533 102 07/27/2005 E05-0210-86550 DLS-SW-TRIP01 < 0.0026 NA 07/27/2005 E05-0210-86551 DLS-SW-TRIP01 LS 0.0534 102 < 0.0026 NA 07/27/2005 E05-0210-86552 DLS-SW-TRIP01 HS 0.508 97.5 07/27/2005 E05-0210-86553 DXS-SW-TRIP02 E05-0210-86554 DXS-SW-TRIP02 LS < 0.0026 0.0530 NA 102 < 0.0026 07/27/2005 NA 07/27/2005 E05-0210-86555 DXS-SW-TRIP02 HS 0.506 97.1 07/27/2005 E05-0210-86556 DXS-SW-TRIP03 < 0.0026 NA 07/27/2005 E05-0210-86557 DXS-SW-TRIP03 LS 0.0528 101 < 0.0026 NA 07/27/2005 E05-0210-86558 DXS-SW-TRIP03 HS 0.531 102 07/27/2005 (1) The analytical uncertainty of the PFHS resuts is 10015.2% based on method accuracy and precision. All results and calculations in this table are presented with three significant figures. Sample concentrations, averages, and %RPD values may vary slightly from the raw data. (2) Low field matrix spike concentration = 0.0521 ng/mL. High field matrix spike concentration = 0.521 ng/mL. (3) NA: Not applicable. %RPD value was not determined when concentrations for the both the sample and sample duplicate were below the stated LOQ. (4) The average value listed is the concentration for the sample or sample duplicate that produced a value above the LOQ. A true average and %RPD between the sample and sample duplicate was not determined. (5) Sample descriptions were misidentified when checked into LIMS. Samples with LIMS ID numbers 86370 and 86371 were in bottles labeled low spike and high spike, respectively. Bottles with LIMS ID numbers 86372 and 86372 were surrogate spiked only (sample and sample duplicate bottles). E05-0210 Interim R eport#13 Tennessee R iver Page 39 of 160 3M Environm ental Labora tory E05-0210 Interim R eport#13 Table 18. PFOS Sample Results. 3M Sample Number Sample Description (1)PFOS Conc. (ng/mL) (2>Field Matrix Spike % Recovery Average Result Sample/Sample Duplicate %RPD Extraction Date E05-0210-86358 DLS-SW-TRM307-0-050718 0.00670 NA 07/26/2005 E05-0210-86359 DLS-SW-TRM307-0-050718 Dup E05-0210-86360 DLS-SW-TRM307-0-050718 LS < 0.00496 0.05960 NA 107 (3)0.00670 07/26/2005 NA 07/26/2005 E05-0210-86361 DLS-SW-TRM307-0-050718 HS 0.564 112 07/26/2005 E05-0210-86362 DLS-SW-TRM301-0-050718 < 0.00496 NA 07/26/2005 E05-0210-86363 DLS-SW-TRM301-0-050718 Dup E05-0210-86364 DLS-SW-TRM301-0-050718 LS < 0.00496 0.0574 NA 116 < 0.00496 (4)n a 07/26/2005 07/26/2005 E05-0210-86365 DLS-SW-TRM301-0-050718 HS 0.607 122 07/26/2005 E05-0210-86366 DLS-SW-TRM301-F-050718 < 0.00496 NA 07/26/2005 E05-0210-86367 DLS-SW-TRM301-F-050718 Dup E05-0210-86368 DLS-SW-TRM301-F-050718 LS < 0.00496 0.0604 NA 122 < 0.00496 (4)n a 07/26/2005 07/26/2005 E05-0210-86369 DLS-SW-TRM301-F-050718 HS 0.558 112 07/26/2005 E05-0210-86370 DLS-SW-TRM295-0-050719(5) 0.12900 111 07/26/2005 E05-0210-86371 DLS-SW-TRM295-0-050719 Dup(5) E05-0210-86372 DLS-SW-TRM295-0-050719 LS(5) 0.59000 0.0736 104 NA 0.0742 07/26/2005 1.48 07/26/2005 E05-0210-86373 DLS-SW-TRM295-0-050719 HS(5) 0.0747 NA 07/26/2005 E05-0210-86374 DLS-SW-TRM289-0-050719 0.07630 NA 07/26/2005 E05-0210-86375 DLS-SW-TRM289-0-050719 Dup E05-0210-86376 DLS-SW-TRM289-0-050719 LS 0.07220 0.1340 NA 121 0.0743 07/26/2005 5.52 07/26/2005 E05-0210-86377 DLS-SW-TRM289-0-050719 HS 0.544 94.7 07/26/2005 E05-0210-86378 DLS-SW-TRM289-F-050719 0.0588 NA 07/26/2005 E05-0210-86379 DLS-SW-TRM289-F-050719 Dup E05-0210-86380 DLS-SW-TRM289-F-050719 LS 0.0462 0.120 NA 136 0.0525 07/26/2005 24.0 07/26/2005 E05-0210-86381 DLS-SW-TRM289-F-050719 HS 0.631 117 07/26/2005 E05-0210-86382 DLS-SW-TRM283-0-050719 0.0528 NA 07/26/2005 E05-0210-86383 DLS-SW-TRM283-0-050719 Dup E05-0210-86384 DLS-SW-TRM283-0-050719 LS 0.0703 0.0973 NA 72.2 0.0616 07/26/2005 28.4 07/26/2005 E05-0210-86385 DLS-SW-TRM283-0-050719 HS 0.605 110 07/26/2005 E05-0210-86386 DLS-SW-TRM277-0-050719 0.0336 NA 07/26/2005 E05-0210-86387 DLS-SW-TRM277-0-050719 Dup E05-0210-86388 DLS-SW-TRM277-0-050719 LS 0.0530 0.0881 NA 90.5 0.0433 44.8 07/26/2005 07/26/2005 E05-0210-86389 DLS-SW-TRM277-0-050719 HS 0.575 107 07/26/2005 E05-0210-86390 DLS-SW-TRM277-F-050719 E05-0210-86391 DLS-SW-TRM277-F-050719 Dup E05-0210-86392 DLS-SW-TRM277-F-050719 LS E05-0210-86393 DLS-SW-TRM277-F-050719 HS 0.0304 0.0390 0.0861 0.515 NA NA 104 96.8 0.0347 07/26/2005 24.8 07/26/2005 07/26/2005 07/26/2005 E05-0210 Interim R eport#13 Tennessee R iver Page 40 of 160 3M Environm ental Labora tory E05-0210 Interim R eport#13 Table 18. PFOS Sample Results Continued. 3M Sample Number Sample Description (1)PFOS Conc. (ng/mL) E05-0210-86394 DLS-SW-TRM271-0-050719 0.0320 (2>Field Matrix Spike % Recovery Average Result Sample/Sample Duplicate %RPD Extraction Date NA 07/26/2005 E05-0210-86395 DLS-SW-TRM271-0-050719 Dup E05-0210-86396 DLS-SW-TRM271-0-050719 LS 0.0332 0.0837 NA 103 0.0326 07/26/2005 3.68 07/26/2005 E05-0210-86397 DLS-SW-TRM271-0-050719 HS 0.563 107 07/26/2005 E05-0210-86398 DLS-SW-TRM265-0-050719 0.0377 NA 07/26/2005 E05-0210-86399 DLS-SW-TRM265-0-050719 Dup E05-0210-86400 DLS-SW-TRM265-0-050719 LS 0.0375 0.0820 NA 89.7 0.0376 0.532 07/26/2005 07/26/2005 E05-0210-86401 DLS-SW-TRM265-0-050719 HS 0.596 112 07/26/2005 E05-0210-86402 DLS-SW-TRM261-2-050720 < 0.00496 NA 07/26/2005 E05-0210-86403 DLS-SW-TRM261-2-050720 Dup E05-0210-86404 DLS-SW-TRM261-2-050720 LS < 0.00496 0.0593 NA 120 < 0.00496 (4)n a 07/26/2005 07/26/2005 E05-0210-86405 DLS-SW-TRM261-2-050720 HS 0.571 115 07/26/2005 E05-0210-86406 DLS-SW-TRM261-0-050720 0.0324 NA 07/26/2005 E05-0210-86407 DLS-SW-TRM261-0-050720 Dup E05-0210-86408 DLS-SW-TRM261-0-050720 LS 0.0330 0.0839 NA 103 0.0327 07/26/2005 1.83 07/26/2005 E05-0210-86409 DLS-SW-TRM261-0-050720 HS 0.562 107 07/26/2005 E05-0210-86410 DLS-SW-TRM261-F-050720 0.0333 NA 07/26/2005 E05-0210-86411 DLS-SW-TRM261-F-050720 Dup E05-0210-86412 DLS-SW-TRM261-F-050720 LS 0.0232 0.0783 NA 101 0.0283 07/26/2005 35.8 07/27/2005 E05-0210-86413 DLS-SW-TRM261-F-050720 HS 0.525 100 07/27/2005 E05-0210-86414 DLS-SW-TRM256-0-050720 0.0326 NA 07/27/2005 E05-0210-86415 DLS-SW-TRM256-0-050720 Dup E05-0210-86416 DLS-SW-TRM256-0-050720 LS 0.0342 0.0856 NA 105 0.0334 4.79 07/27/2005 07/27/2005 E05-0210-86417 DLS-SW-TRM256-0-050720 HS 0.558 106 07/27/2005 E05-0210-86418 DLS-SW-TRM254-0-050720 0.0352 NA 07/27/2005 E05-0210-86419 DLS-SW-TRM254-0-050720 Dup E05-0210-86420 DLS-SW-TRM254-0-050720 LS 0.0341 0.0927 NA 117 0.0347 07/27/2005 3.17 07/27/2005 E05-0210-86421 DLS-SW-TRM254-0-050720 HS 0.594 113 07/27/2005 E05-0210-86422 DLS-SW-TRM254-F-050720 0.0300 NA 07/27/2005 E05-0210-86423 DLS-SW-TRM254-F-050720 Dup E05-0210-86424 DLS-SW-TRM254-F-050720 LS 0.0342 0.0903 NA 118 0.0321 07/27/2005 13.1 07/27/2005 E05-0210-86425 DLS-SW-TRM254-F-050720 HS 0.626 120 07/27/2005 E05-0210-86426 DLS-SW-TRM254-2-050720 < 0.00992 NA 07/27/2005 E05-0210-86427 DLS-SW-TRM254-2-050720 Dup E05-0210-86428 DLS-SW-TRM254-2-050720 LS < 0.00992 0.0577 NA 116 < 0.00992 (4)n a 07/27/2005 07/27/2005 E05-0210-86429 DLS-SW-TRM254-2-050720 HS 0.525 106 07/27/2005 E05-0210 Interim R eport#13 Tennessee R iver Page 41 of 160 3M Environm ental Labora tory E05-0210 Interim R eport#13 Table 18. PFOS Sample Results Continued. 3M Sample Number Sample Description E05-0210-86430 DXS-SW-T01001-0-050721 E05-0210-86431 DXS-SW-T01001-0-050721 Dup E05-0210-86432 DXS-SW-T01001-0-050721 LS E05-0210-86433 DXS-SW-T01001-0-050721 HS E05-0210-86434 DXS-SW-T01002-0-050721 E05-0210-86435 DXS-SW-T01002-0-050721 Dup E05-0210-86436 DXS-SW-T01002-0-050721 LS E05-0210-86437 DXS-SW-T01002-0-050721 HS E05-0210-86438 DXS-SW-T01003-0-050721 E05-0210-86439 DXS-SW-T01003-0-050721 Dup E05-0210-86440 DXS-SW-T01003-0-050721 LS E05-0210-86441 DXS-SW-T01003-0-050721 HS E05-0210-86442 DXS-SW-T01004-0-050721 E05-0210-86443 DXS-SW-T01004-0-050721 Dup E05-0210-86444 DXS-SW-T01004-0-050721 LS E05-0210-86445 DXS-SW-T01004-0-050721 HS E05-0210-86446 DXS-SW-T01005-0-050721 E05-0210-86447 DXS-SW-T01005-0-050721 Dup E05-0210-86448 DXS-SW-T01005-0-050721 LS E05-0210-86449 DXS-SW-T01005-0-050721 HS E05-0210-86450 DXS-SW-T01006-0-050721 E05-0210-86451 DXS-SW-T01006-0-050721 Dup E05-0210-86452 DXS-SW-T01006-0-050721 LS E05-0210-86453 DXS-SW-T01006-0-050721 HS E05-0210-86454 DXS-SW-T01007-0-050721 E05-0210-86455 DXS-SW-T01007-0-050721 Dup E05-0210-86456 DXS-SW-T01007-0-050721 LS E05-0210-86457 DXS-SW-T01007-0-050721 HS E05-0210-86458 DXS-SW-T01008-0-050721 E05-0210-86459 DXS-SW-T01008-0-050721 Dup E05-0210-86460 DXS-SW-T01008-0-050721 LS E05-0210-86461 DXS-SW-T01008-0-050721 HS E05-0210-86462 DXS-SW-T01009-0-050721 E05-0210-86463 DXS-SW-T01009-0-050721 Dup E05-0210-86464 DXS-SW-T01009-0-050721 LS E05-0210-86465 DXS-SW-T01009-0-050721 HS (1)PFOS Conc. (ng/mL) 0.0739 0.0680 0.1230 0.602 0.0607 0.0594 0.1050 0.635 0.0542 0.0555 0.107 0.580 0.0560 0.0636 0.114 0.521 0.0383 0.0390 0.133 0.605 0.0209 0.0219 0.0903 0.650 0.0273 0.0268 0.0865 0.586 0.0219 0.0217 0.0892 0.523 0.0202 0.0203 0.0770 0.631 Average (2>Field Matrix Result Spike % Sample/Sample Recovery Duplicate %RPD Extraction Date NA 07/27/2005 NA 07/27/2005 0.0710 8.32 105 07/27/2005 107 07/27/2005 NA 07/27/2005 NA 90.8 0.0601 07/27/2005 2.16 07/27/2005 116 07/27/2005 NA 07/27/2005 NA 07/27/2005 105 0.0549 2.37 07/27/2005 117 07/28/2005 centrifuged NA 07/27/2005 NA 07/27/2005 0.0598 12.7 109 07/27/2005 93.0 07/27/2005 NA 07/27/2005 NA 07/27/2005 0.0387 1.81 191 07/27/2005 114 07/27/2005 NA 07/27/2005 NA 07/27/2005 0.0214 4.67 139 07/27/2005 127 07/27/2005 NA 07/27/2005 NA 07/27/2005 0.0271 1.85 120 07/27/2005 113 07/27/2005 NA 07/27/2005 NA 07/27/2005 0.0218 0.917 136 07/27/2005 101 07/27/2005 NA 07/27/2005 NA 07/27/2005 0.0203 0.494 115 07/27/2005 123 07/27/2005 E05-0210 Interim R eport#13 Tennessee R iver Page 42 of 160 3M Environm ental Laboratory E05-0210 Interim R eport#13 Table 18. PFOS Sample Results Continued. 3M Sample Number Sample Description (1)PFOS Conc. (ng/mL) Average <2>Field Matrix Result Spike % Sample/Sample Recovery Duplicate %RPD Extraction Date E05-0210-86466 DXS-SW-T01010-0-050721 0.0216 NA 07/27/2005 E05-0210-86467 DXS-SW-T01010-0-050721 Dup E05-0210-86468 DXS-SW-T01010-0-050721 LS 0.0219 0.0772 NA 112 0.0218 07/27/2005 1.38 07/27/2005 E05-0210-86469 DXS-SW-T01010-0-050721 HS 0.553 107 07/27/2005 E05-0210-86470 DXS-SW-T02001-0-050721 0.129 NA 07/27/2005 E05-0210-86471 DXS-SW-T02001-0-050721 Dup E05-0210-86472 DXS-SW-T02001-0-050721 LS 0.117 0.170 NA 94.9 0.123 07/27/2005 9.76 07/27/2005 E05-0210-86473 DXS-SW-T02001-0-050721 HS 0.640 104 07/27/2005 E05-0210-86474 DXS-SW-T02002-0-050721 0.0707 NA 07/27/2005 E05-0210-86475 DXS-SW-T02002-0-050721 Dup E05-0210-86476 DXS-SW-T02002-0-050721 LS 0.0762 0.1260 NA 106 0.0735 07/27/2005 7.49 07/27/2005 E05-0210-86477 DXS-SW-T02002-0-050721 HS 0.606 107 07/27/2005 E05-0210-86478 DXS-SW-T02003-0-050721 < 0.00992 NA 07/27/2005 E05-0210-86479 DXS-SW-T02003-0-050721 Dup E05-0210-86480 DXS-SW-T02003-0-050721 LS < 0.00992 0.0548 NA 111 < 0.00992 (4)n a 07/27/2005 07/27/2005 E05-0210-86481 DXS-SW-T02003-0-050721 HS 0.511 103 07/27/2005 E05-0210-86482 DXS-SW-T02004-0-050721 < 0.00992 NA 07/27/2005 E05-0210-86483 DXS-SW-T02004-0-050721 Dup E05-0210-86484 DXS-SW-T02004-0-050721 LS < 0.00992 0.0659 NA 119 < 0.00992 07/27/2005 NA 07/27/2005 E05-0210-86485 DXS-SW-T02004-0-050721 HS 0.523 104 07/27/2005 E05-0210-86486 DXS-SW-T02005-0-050721 < 0.00744 NA 07/27/2005 E05-0210-86487 DXS-SW-T02005-0-050721 Dup E05-0210-86488 DXS-SW-T02005-0-050721 LS < 0.0496 0.0560 NA 113 < 0.0496 (4)n a 07/28/2005 c e n trifu g e d 07/27/2005 E05-0210-86489 DXS-SW-T02005-0-050721 HS 0.684 138 07/28/2005 c e n trifu g e d E05-0210-86490 DXS-SW-T02006-0-050721 < 0.00744 NA 07/27/2005 E05-0210-86491 DXS-SW-T02006-0-050721 Dup E05-0210-86492 DXS-SW-T02006-0-050721 LS 0.00799 0.0728 NA 131 (3)(6)0.00799 07/27/2005 NA 07/27/2005 E05-0210-86493 DXS-SW-T02006-0-050721 HS 0.667 133 07/27/2005 E05-0210-86494 DXS-SW-T02007-0-050721 < 0.00744 NA 07/27/2005 E05-0210-86495 DXS-SW-T02007-0-050721 Dup E05-0210-86496 DXS-SW-T02007-0-050721 LS < 0.00744 0.0513 NA 104 < 0.00744 (4)n a 07/27/2005 07/27/2005 E05-0210-86497 DXS-SW-T02007-0-050721 HS 0.479 96.6 07/27/2005 E05-0210-86498 DXS-SW-T02008-0-050721 < 0.00744 NA 07/27/2005 E05-0210-86499 DXS-SW-T02008-0-050721 Dup E05-0210-86500 DXS-SW-T02008-0-050721 LS < 0.00744 0.0579 NA 117 < 0.00744 (4)n a 07/27/2005 07/27/2005 E05-0210-86501 DXS-SW-T02008-0-050721 HS 0.493 99.4 07/27/2005 E05-0210 Interim R eport#13 Tennessee R iver Page 43 of 160 3M Environm ental Labora tory E05-0210 Interim R eport#13 Table 18. PFOS Sample Results Continued. 3M Sample Number Sample Description (1)PFOS Conc. (ng/mL) Average (2>Field Matrix Result Spike % Sample/Sample Recovery Duplicate %RPD Extraction Date E05-0210-86502 DXS-SW-T02009-0-050721 0.00872 NA 07/27/2005 E05-0210-86503 DXS-SW-T02009-0-050721 Dup E05-0210-86504 DXS-SW-T02009-0-050721 LS 0.00883 0.0725 NA 129 0.00878 07/27/2005 1.25 07/27/2005 E05-0210-86505 DXS-SW-T02009-0-050721 HS 0.524 104 07/27/2005 E05-0210-86506 DXS-SW-T02010-0-050721 0.0426 NA 07/27/2005 E05-0210-86507 DXS-SW-T02010-0-050721 Dup E05-0210-86508 DXS-SW-T02010-0-050721 LS 0.0285 0.0590 NA 47.4 (6)0.0356 07/27/2005 39.7 07/27/2005 E05-0210-86509 DXS-SW-T02010-0-050721 HS 0.293 51.9 07/27/2005 E05-0210-86510 DXS-SW-T03001-0-050721 0.782 NA 07/27/2005 E05-0210-86511 DXS-SW-T03001-0-050721 Dup E05-0210-86512 DXS-SW-T03001-0-050721 LS 0.807 0.909 NA (7)n r 0.795 07/27/2005 3.15 07/27/2005 E05-0210-86513 DXS-SW-T03001-0-050721 HS 1.330 108 07/27/2005 E05-0210-86514 DXS-SW-T03002-0-050721 0.0235 NA 07/27/2005 E05-0210-86515 DXS-SW-T03002-0-050721 Dup E05-0210-86516 DXS-SW-T03002-0-050721 LS 0.0169 0.0711 NA 103 0.0202 07/27/2005 32.7 07/27/2005 E05-0210-86517 DXS-SW-T03002-0-050721 HS 0.543 105 07/27/2005 E05-0210-86518 DXS-SW-T03003-0-050721 0.0111 NA 07/27/2005 E05-0210-86519 DXS-SW-T03003-0-050721 Dup E05-0210-86520 DXS-SW-T03003-0-050721 LS 0.0118 0.0715 NA 121 0.0115 07/27/2005 6.11 07/27/2005 E05-0210-86521 DXS-SW-T03003-0-050721 HS 0.549 108 07/27/2005 E05-0210-86522 DXS-SW-T03004-0-050722 0.0226 NA 07/27/2005 E05-0210-86523 DXS-SW-T03004-0-050722 Dup E05-0210-86524 DXS-SW-T03004-0-050722 LS 0.0167 0.0696 NA 101 0.0197 07/27/2005 30.0 07/27/2005 E05-0210-86525 DXS-SW-T03004-0-050722 HS 0.561 109 07/27/2005 E05-0210-86526 DXS-SW-T03005-0-050722 < 0.00744 NA 07/27/2005 E05-0210-86527 DXS-SW-T03005-0-050722 Dup E05-0210-86528 DXS-SW-T03005-0-050722 LS 0.0773 0.0762 NA (8)n r (3)0.0773 07/28/2005 NA c e n trifu g e d 07/27/2005 E05-0210-86529 DXS-SW-T03005-0-050722 HS 0.594 104 07/27/2005 E05-0210-86530 DXS-SW-T03006-0-050722 0.00763 NA 07/27/2005 E05-0210-86531 DXS-SW-T03006-0-050722 Dup E05-0210-86532 DXS-SW-T03006-0-050722 LS E05-0210-86533 DXS-SW-T03006-0-050722 HS < 0.00744 < 0.0496 0.665 NA (9)n r 132 (3)0.00763 07/27/2005 NA 07/28/2005 c e n trifu g e d 07/27/2005 E05-0210-86534 DXS-SW-T03007-0-050722 < 0.00744 NA 07/27/2005 E05-0210-86535 DXS-SW-T03007-0-050722 Dup E05-0210-86536 DXS-SW-T03007-0-050722 LS E05-0210-86537 DXS-SW-T03007-0-050722 HS < 0.0496 0.0664 0.576 NA 134 116 < 0.0496 (4)n a 07/28/2005 c e n trifu g e d 07/27/2005 07/27/2005 E05-0210 Interim R eport#13 Tennessee R iver Page 44 of 160 3M Environm ental Laboratory E05-0210 Interim R eport#13 Table 18. PFOS Sample Results Continued. 3M Sample Number Sample Description (1)PFOS Conc. (ng/mL) Average <2>Field Matrix Result Spike % Sample/Sample Recovery Duplicate %RPD Extraction Date E05-0210-86538 DXS-SW-T03008-0-050722 < 0.00744 NA 07/27/2005 E05-0210-86539 DXS-SW-T03008-0-050722 Dup E05-0210-86540 DXS-SW-T03008-0-050722 LS < 0.00744 0.0600 NA 121 < 0.00744 07/27/2005 NA 07/27/2005 E05-0210-86541 DXS-SW-T03008-0-050722 HS 0.561 113 07/27/2005 E05-0210-86542 DXS-SW-T03009-0-050722 0.00809 NA 07/27/2005 E05-0210-86543 DXS-SW-T03009-0-050722 Dup E05-0210-86544 DXS-SW-T03009-0-050722 LS < 0.00744 0.0608 NA 106 (3)0.00809 07/27/2005 NA 07/27/2005 E05-0210-86545 DXS-SW-T03009-0-050722 HS 0.508 101 07/27/2005 E05-0210-86546 DXS-SW-T03010-0-050722 0.00749 NA 07/27/2005 E05-0210-86547 DXS-SW-T03010-0-050722 Dup E05-0210-86548 DXS-SW-T03010-0-050722 LS < 0.00744 0.0556 NA 97.2 (3)0.00749 07/27/2005 NA 07/27/2005 E05-0210-86549 DXS-SW-T03010-0-050722 HS 0.506 100 07/27/2005 E05-0210-86550 DLS-SW-TRIP01 E05-0210-86551 DLS-SW-TRIP01 LS < 0.00744 0.0551 NA 111 < 0.00744 07/27/2005 NA 07/27/2005 E05-0210-86552 DLS-SW-TRIP01 HS 0.457 92.1 07/27/2005 E05-0210-86553 DXS-SW-TRIP02 < 0.00744 NA 07/27/2005 E05-0210-86554 DXS-SW-TRIP02 LS 0.0516 104 < 0.00744 NA 07/27/2005 E05-0210-86555 DXS-SW-TRIP02 HS 0.491 99.0 07/27/2005 E05-0210-86556 DXS-SW-TRIP03 < 0.00744 NA 07/27/2005 E05-0210-86557 DXS-SW-TRIP03 LS 0.0552 112 < 0.00744 NA 07/27/2005 E05-0210-86558 DXS-SW-TRIP03 HS 0.537 108 07/27/2005 (1) The analytical uncertainty of the PFOS results is 10013.8 % based on method accuracy and precision. All results and calculations in this table are presented with three significant figures. Sample concentrations, averages, and %RPD values may vary slightly from the raw data. (2) Low field matrix spike concentration = 0.0496 ng/mL. High field matrix spike concentration = 0.496 ng/mL. (3) The average value listed is the concentration for the sample or sample duplicate that produced a value above the LOQ. A true average and %RPD between the sample and sample duplicate was not determined. (4) NA: Not applicable. %RPD value was not determined when concentrations for the both the sample and sample duplicate were below the stated LOQ. (5) Sample descriptions were misidentified when checked into LIMS. Samples with LIMS ID numbers 86370 and 86371 were in bottles labeled low spike and high spike, respectively. Bottles with LIMS ID numbers 86372 and 86372 were surrogate spiked only (sample and sample duplicate bottles). (6) Both field matrix spikes associated with this sampling location exceeded method acceptance criteria of 10030%. Analytical accuracy for this location should be considered 10050% due matrix contributions. (7) NR: Not reported. Spike recovery not calculated as the endogenous sample concentration was at least three times the spiked amount. (8) NR: Not reported. Concentration of spiked sample roughly equivalent to sample replicate producing a concentration above the stated LOQ. Reported sample concentration should be considered accurate to within 10050% based on high field matrix spike recovery only (poor precision between sample/sample duplicate). (9) NR: Not reported. PFOS not quantitated above the stated LOQ for the low matrix spike. Reported sample concentration should be considered accurate to within 10050% based on high field matrix spike recovery only. E05-0210 Interim R eport#13 Tennessee R iver Page 45 of 160 Table 19. PFOA[1,2 13C] Surrogate Results. 3M Environm ental Labora tory E05-0210 Interim R eport#13 3M Sample Number E05-0210-86358 E05-0210-86359 E05-0210-86360 E05-0210-86361 E05-0210-86362 E05-0210-86363 E05-0210-86364 E05-0210-86365 E05-0210-86366 E05-0210-86367 E05-0210-86368 E05-0210-86369 E05-0210-86370 E05-0210-86371 E05-0210-86372 E05-0210-86373 E05-0210-86374 E05-0210-86375 E05-0210-86376 E05-0210-86377 E05-0210-86378 E05-0210-86379 E05-0210-86380 E05-0210-86381 E05-0210-86382 E05-0210-86383 E05-0210-86384 E05-0210-86385 E05-0210-86386 E05-0210-86387 E05-0210-86388 E05-0210-86389 E05-0210-86390 E05-0210-86391 E05-0210-86392 E05-0210-86393 E05-0210-86394 E05-0210-86395 E05-0210-86396 E05-0210-86397 Sample Description DLS-SW-TRM307-0-050718 DLS-SW-TRM307-0-050718 Dup DLS-SW-TRM307-0-050718 LS DLS-SW-TRM307-0-050718 HS DLS-SW-TRM301 -0-050718 DLS-SW-TRM301-0-050718 Dup DLS-SW-TRM301-0-050718 LS DLS-SW-TRM301-0-050718 HS DLS-SW-TRM301 -F-050718 DLS-SW-TRM301-F-050718 Dup DLS-SW-TRM301-F-050718 LS DLS-SW-TRM301-F-050718 HS DLS-SW-TRM295-0-050719 DLS-SW-TRM295-0-050719 Dup DLS-SW-TRM295-0-050719 LS DLS-SW-TRM295-0-050719 HS DLS-SW-TRM289-0-050719 DLS-SW-TRM289-0-050719 Dup DLS-SW-TRM289-0-050719 LS DLS-SW-TRM289-0-050719 HS DLS-SW-TRM289-F-050719 DLS-SW-TRM289-F-050719 Dup DLS-SW-TRM289-F-050719 LS DLS-SW-TRM289-F-050719 HS DLS-SW-TRM283-0-050719 DLS-SW-TRM283-0-050719 Dup DLS-SW-TRM283-0-050719 LS DLS-SW-TRM283-0-050719 HS DLS-SW-TRM277-0-050719 DLS-SW-TRM277-0-050719 Dup DLS-SW-TRM277-0-050719 LS DLS-SW-TRM277-0-050719 HS DLS-SW-TRM277-F-050719 DLS-SW-TRM277-F-050719 Dup DLS-SW-TRM277-F-050719 LS DLS-SW-TRM277-F-050719 HS DLS-SW-TRM271 -0-050719 DLS-SW-TRM271-0-050719 Dup DLS-SW-TRM271-0-050719 LS DLS-SW-TRM271-0-050719 HS (2>Non-Corrected (3>Corrected (1>PFOA [1,2 13C] Surrogate Surrogate Concentration Percent Percent (ng/mL> Recovery Recovery Extraction Date 0.177 0.162 0.160 0.154 64.6 59.1 63.0 60.6 115 07/26/2005 105 07/26/2005 112 07/26/2005 108 07/26/2005 0.155 0.155 0.146 0.148 56.6 56.6 57.5 58.3 101 07/26/2005 101 07/26/2005 102 07/26/2005 104 07/26/2005 0.160 0.172 0.153 0.146 58.4 62.8 60.2 57.5 104 07/26/2005 112 07/26/2005 107 07/26/2005 102 07/26/2005 0.158 0.137 0.152 0.158 62.2 53.9 55.5 57.7 111 07/26/2005 96.1 07/26/2005 98.9 07/26/2005 103 07/26/2005 0.156 0.154 0.142 0.131 56.9 56.2 55.9 51.6 101 07/26/2005 100 07/26/2005 99.6 07/26/2005 91.9 07/26/2005 0.150 0.165 0.154 0.145 54.7 60.2 60.6 57.1 97.6 07/26/2005 107 07/26/2005 108 07/26/2005 102 07/26/2005 0.164 0.154 0.149 0.146 59.8 56.2 58.7 57.5 107 07/26/2005 100 07/26/2005 104 07/26/2005 102 07/26/2005 0.160 0.153 0.159 0.133 58.4 55.8 62.6 52.4 104 07/26/2005 99.5 07/26/2005 112 07/26/2005 93.3 07/26/2005 0.161 0.169 0.157 0.122 58.8 61.7 61.8 48.0 105 07/26/2005 110 07/26/2005 110 07/26/2005 85.6 07/26/2005 0.153 0.158 0.135 0.136 55.8 57.7 53.1 53.5 99.5 103 94.7 95.4 07/26/2005 07/26/2005 07/26/2005 07/26/2005 E05-0210 Interim R eport#13 Tennessee R iver Page 46 of 160 3M Environm ental Labora tory E05-0210 Interim R eport#13 Table 19. PFOA[1,2 13C] Surrogate Results Continued. 3M Sample Number Sample Description (2)Non-Corrected (3)Corrected (1)PFOA [1,2 13C] Surrogate Surrogate Concentration Percent Percent (ng/mL) Recovery Recovery Extraction Date E05-0210-86398 E05-0210-86399 E05-0210-86400 E05-0210-86401 DLS-SW-TRM265-0-050719 DLS-SW-TRM265-0-050719 Dup DLS-SW-TRM265-0-050719 LS DLS-SW-TRM265-0-050719 HS 0.148 0.157 0.133 0.138 54.0 57.3 52.4 54.3 96.3 07/26/2005 102 07/26/2005 93.3 07/26/2005 96.8 07/26/2005 E05-0210-86402 E05-0210-86403 E05-0210-86404 E05-0210-86405 DLS-SW-TRM261 -2-050720 DLS-SW-TRM261-2-050720 Dup DLS-SW-TRM261-2-050720 LS DLS-SW-TRM261-2-050720 HS 0.158 0.159 0.150 0.129 57.7 58.0 59.0 50.8 103 07/26/2005 103 07/26/2005 105 07/26/2005 90.5 07/26/2005 E05-0210-86406 E05-0210-86407 E05-0210-86408 E05-0210-86409 DLS-SW-TRM261 -0-050720 DLS-SW-TRM261-0-050720 Dup DLS-SW-TRM261-0-050720 LS DLS-SW-TRM261-0-050720 HS 0.147 0.156 0.141 0.134 53.6 56.9 55.5 52.8 95.6 07/26/2005 101 07/26/2005 99.0 07/26/2005 94.0 07/26/2005 E05-0210-86410 E05-0210-86411 E05-0210-86412 E05-0210-86413 DLS-SW-TRM261 -F-050720 DLS-SW-TRM261 -F-050720 Dup DLS-SW-TRM261-F-050720 LS DLS-SW-TRM261-F-050720 HS 0.154 0.141 0.140 0.123 56.2 51.4 55.1 48.4 100 07/26/2005 91.7 07/26/2005 98.2 07/27/2005 86.3 07/27/2005 E05-0210-86414 E05-0210-86415 E05-0210-86416 E05-0210-86417 DLS-SW-TRM256-0-050720 DLS-SW-TRM256-0-050720 Dup DLS-SW-TRM256-0-050720 LS DLS-SW-TRM256-0-050720 HS 0.143 0.148 0.145 0.133 52.2 54.0 57.1 52.4 93.0 07/27/2005 96.3 07/27/2005 102 07/27/2005 93.3 07/27/2005 E05-0210-86418 E05-0210-86419 E05-0210-86420 E05-0210-86421 DLS-SW-TRM254-0-050720 DLS-SW-TRM254-0-050720 Dup DLS-SW-TRM254-0-050720 LS DLS-SW-TRM254-0-050720 HS 0.158 0.154 0.158 0.132 57.7 56.2 62.2 52.0 103 07/27/2005 100 07/27/2005 111 07/27/2005 92.6 07/27/2005 E05-0210-86422 E05-0210-86423 E05-0210-86424 E05-0210-86425 DLS-SW-TRM254-F-050720 DLS-SW-TRM254-F-050720 Dup DLS-SW-TRM254-F-050720 LS DLS-SW-TRM254-F-050720 HS 0.160 0.164 0.159 0.147 58.4 59.8 62.6 57.9 104 07/27/2005 107 07/27/2005 112 07/27/2005 103 07/27/2005 E05-0210-86426 E05-0210-86427 E05-0210-86428 E05-0210-86429 DLS-SW-TRM254-2-050720 DLS-SW-TRM254-2-050720 Dup DLS-SW-TRM254-2-050720 LS DLS-SW-TRM254-2-050720 HS 0.162 0.157 0.145 0.138 59.1 57.3 57.1 54.3 105 07/27/2005 102 07/27/2005 102 07/27/2005 96.8 07/27/2005 E05-0210-86430 E05-0210-86431 E05-0210-86432 E05-0210-86433 DXS-SW-T01001 -0-050721 DXS-SW-T01001-0-050721 Dup DXS-SW-T01001-0-050721 LS DXS-SW-T01001-0-050721 HS 0.172 0.164 0.159 0.142 62.8 59.8 62.6 55.9 112 07/27/2005 107 07/27/2005 112 07/27/2005 99.6 07/27/2005 E05-0210-86434 E05-0210-86435 E05-0210-86436 E05-0210-86437 DXS-SW-T01002-0-050721 DXS-SW-T01002-0-050721 Dup DXS-SW-T01002-0-050721 LS DXS-SW-T01002-0-050721 HS 0.167 0.164 0.139 0.146 60.9 59.8 54.7 57.5 109 07/27/2005 107 07/27/2005 97.6 07/27/2005 102 07/27/2005 E05-0210 Interim R eport#13 Tennessee R iver Page 47 of 160 3M Environm ental Labora tory E05-0210 Interim R eport#13 Table 19. PFOA[1,2 13C] Surrogate Results Continued. 3 M Sam ple N um ber Sam ple Description (2)N o n -C o rre c te d (3)C o rrec te d (1)P F O A [1 ,2 13C ] S u rro g a te Surrogate C o n c e n tra tio n Percent Percent (ng/m L) Recovery Recovery E x tra c tio n D ate E 0 5 -0 2 1 0 -8 6 4 3 8 E 0 5 -0 2 1 0 -8 6 4 3 9 E 0 5 -0 2 1 0 -8 6 4 4 0 E 0 5 -0 2 1 0 -8 6 4 4 1 E 0 5 -0 2 1 0 -8 6 4 4 2 E 0 5 -0 2 1 0 -8 6 4 4 3 E 0 5 -0 2 1 0 -8 6 4 4 4 E 0 5 -0 2 1 0 -8 6 4 4 5 D X S -S W -T 0 1 0 0 3 -0 -0 5 0 7 2 1 D X S -S W -T 0 1 0 0 3 -0 -0 5 0 7 2 1 D u p D X S -S W -T 0 1 0 0 3 -0 -0 5 0 7 2 1 LS D X S -S W -T 0 1 0 0 3 -0 -0 5 0 7 2 1 H S D X S -S W -T 0 1 0 0 4 -0 -0 5 0 7 2 1 D X S -S W -T 0 1 0 0 4 -0 -0 5 0 7 2 1 D u p D X S -S W -T 0 1 0 0 4 -0 -0 5 0 7 2 1 LS D X S -S W -T 0 1 0 0 4 -0 -0 5 0 7 2 1 H S 0 .1 5 8 0 .1 6 1 0 .1 4 2 0 .1 3 5 0 .1 7 5 0 .1 6 0 0 .1 6 2 0 .1 2 6 5 7 .7 5 8 .8 5 5 .9 5 3 .1 6 3 .9 5 8 .4 6 3 .8 4 9 .6 103 105 9 9 .6 9 4 .7 114 104 114 8 8 .4 0 7 /2 7 /2 0 0 5 0 7 /2 7 /2 0 0 5 0 7 /2 7 /2 0 0 5 0 7 /2 8 /2 0 0 5 c e n trifu g e d 0 7 /2 7 /2 0 0 5 0 7 /2 7 /2 0 0 5 0 7 /2 7 /2 0 0 5 0 7 /2 7 /2 0 0 5 E 0 5 -0 2 1 0 -8 6 4 4 6 E 0 5 -0 2 1 0 -8 6 4 4 7 E 0 5 -0 2 1 0 -8 6 4 4 8 E 0 5 -0 2 1 0 -8 6 4 4 9 D X S -S W -T 0 1 0 0 5 -0 -0 5 0 7 2 1 D X S -S W -T 0 1 0 0 5 -0 -0 5 0 7 2 1 D u p D X S -S W -T 0 1 0 0 5 -0 -0 5 0 7 2 1 LS D X S -S W -T 0 1 0 0 5 -0 -0 5 0 7 2 1 H S 0 .1 6 7 0 .1 7 1 0 .1 5 1 0 .1 3 7 6 0 .9 6 2 .4 5 9 .4 5 3 .9 109 111 106 9 6 .1 0 7 /2 7 /2 0 0 5 0 7 /2 7 /2 0 0 5 0 7 /2 7 /2 0 0 5 0 7 /2 7 /2 0 0 5 E 0 5 -0 2 1 0 -8 6 4 5 0 E 0 5 -0 2 1 0 -8 6 4 5 1 E 0 5 -0 2 1 0 -8 6 4 5 2 E 0 5 -0 2 1 0 -8 6 4 5 3 D X S -S W -T 0 1 0 0 6 -0 -0 5 0 7 2 1 D X S -S W -T 0 1 0 0 6 -0 -0 5 0 7 2 1 D u p D X S -S W -T 0 1 0 0 6 -0 -0 5 0 7 2 1 LS D X S -S W -T 0 1 0 0 6 -0 -0 5 0 7 2 1 H S 0 .1 5 7 0 .1 6 2 0 .1 7 8 0 .1 5 1 5 7 .3 5 9 .1 7 0 .1 5 9 .4 102 105 125 106 0 7 /2 7 /2 0 0 5 0 7 /2 7 /2 0 0 5 0 7 /2 7 /2 0 0 5 0 7 /2 7 /2 0 0 5 E 0 5 -0 2 1 0 -8 6 4 5 4 E 0 5 -0 2 1 0 -8 6 4 5 5 E 0 5 -0 2 1 0 -8 6 4 5 6 E 0 5 -0 2 1 0 -8 6 4 5 7 D X S -S W -T 0 1 0 0 7 -0 -0 5 0 7 2 1 D X S -S W -T 0 1 0 0 7 -0 -0 5 0 7 2 1 D u p D X S -S W -T 0 1 0 0 7 -0 -0 5 0 7 2 1 LS D X S -S W -T 0 1 0 0 7 -0 -0 5 0 7 2 1 H S 0 .1 4 8 0 .1 5 9 0 .1 5 2 0 .1 4 4 5 4 .0 5 8 .0 5 9 .8 5 6 .7 9 6 .3 103 107 101 0 7 /2 7 /2 0 0 5 0 7 /2 7 /2 0 0 5 0 7 /2 7 /2 0 0 5 0 7 /2 7 /2 0 0 5 E 0 5 -0 2 1 0 -8 6 4 5 8 E 0 5 -0 2 1 0 -8 6 4 5 9 E 0 5 -0 2 1 0 -8 6 4 6 0 E 0 5 -0 2 1 0 -8 6 4 6 1 D X S -S W -T 0 1 0 0 8 -0 -0 5 0 7 2 1 D X S -S W -T 0 1 0 0 8 -0 -0 5 0 7 2 1 D u p D X S -S W -T 0 1 0 0 8 -0 -0 5 0 7 2 1 LS D X S -S W -T 0 1 0 0 8 -0 -0 5 0 7 2 1 H S 0 .1 4 9 0 .1 4 9 0 .1 8 0 0 .1 2 3 5 4 .4 5 4 .4 7 0 .9 4 8 .4 9 6 .9 9 6 .9 126 8 6 .3 0 7 /2 7 /2 0 0 5 0 7 /2 7 /2 0 0 5 0 7 /2 7 /2 0 0 5 0 7 /2 7 /2 0 0 5 E 0 5 -0 2 1 0 -8 6 4 6 2 E 0 5 -0 2 1 0 -8 6 4 6 3 E 0 5 -0 2 1 0 -8 6 4 6 4 E 0 5 -0 2 1 0 -8 6 4 6 5 D X S -S W -T 0 1 0 0 9 -0 -0 5 0 7 2 1 D X S -S W -T 0 1 0 0 9 -0 -0 5 0 7 2 1 D u p D X S -S W -T 0 1 0 0 9 -0 -0 5 0 7 2 1 LS D X S -S W -T 0 1 0 0 9 -0 -0 5 0 7 2 1 H S 0 .1 6 3 0 .1 6 3 0 .1 4 4 0 .1 4 1 5 9 .5 5 9 .5 5 6 .7 5 5 .5 106 106 101 9 9 .0 0 7 /2 7 /2 0 0 5 0 7 /2 7 /2 0 0 5 0 7 /2 7 /2 0 0 5 0 7 /2 7 /2 0 0 5 E 0 5 -0 2 1 0 -8 6 4 6 6 E 0 5 -0 2 1 0 -8 6 4 6 7 E 0 5 -0 2 1 0 -8 6 4 6 8 E 0 5 -0 2 1 0 -8 6 4 6 9 D X S -S W -T 0 1 0 1 0 -0 -0 5 0 7 2 1 D X S -S W -T 0 1 0 1 0 -0 -0 5 0 7 2 1 D u p D X S -S W -T 0 1 0 1 0 -0 -0 5 0 7 2 1 LS D X S -S W -T 0 1 0 1 0 -0 -0 5 0 7 2 1 H S 0 .1 6 8 0 .1 7 0 0 .1 4 5 0 .1 3 4 6 1 .3 6 2 .0 5 7 .1 5 2 .8 109 110 102 9 4 .0 0 7 /2 7 /2 0 0 5 0 7 /2 7 /2 0 0 5 0 7 /2 7 /2 0 0 5 0 7 /2 7 /2 0 0 5 E 0 5 -0 2 1 0 -8 6 4 7 0 E 0 5 -0 2 1 0 -8 6 4 7 1 E 0 5 -0 2 1 0 -8 6 4 7 2 E 0 5 -0 2 1 0 -8 6 4 7 3 D X S -S W -T 0 2 0 0 1 -0 -0 5 0 7 2 1 D X S -S W -T 0 2 0 0 1 -0 -0 5 0 7 2 1 D u p D X S -S W -T 0 2 0 0 1 -0 -0 5 0 7 2 1 LS D X S -S W -T 0 2 0 0 1 -0 -0 5 0 7 2 1 H S 0 .1 5 2 0 .1 6 5 0 .1 4 1 0 .1 3 1 5 5 .5 6 0 .2 5 5 .5 5 1 .6 9 8 .9 107 9 9 .0 9 1 .9 0 7 /2 7 /2 0 0 5 0 7 /2 7 /2 0 0 5 0 7 /2 7 /2 0 0 5 0 7 /2 7 /2 0 0 5 E 0 5 -0 2 1 0 -8 6 4 7 4 E 0 5 -0 2 1 0 -8 6 4 7 5 E 0 5 -0 2 1 0 -8 6 4 7 6 E 0 5 -0 2 1 0 -8 6 4 7 7 D X S -S W -T 0 2 0 0 2 -0 -0 5 0 7 2 1 D X S -S W -T 0 2 0 0 2 -0 -0 5 0 7 2 1 D u p D X S -S W -T 0 2 0 0 2 -0 -0 5 0 7 2 1 LS D X S -S W -T 0 2 0 0 2 -0 -0 5 0 7 2 1 H S 0 .1 4 8 0 .1 4 5 0 .1 3 8 0 .1 3 7 5 4 .0 5 2 .9 5 4 .3 5 3 .9 9 6 .3 9 4 .3 9 6 .8 9 6 .1 0 7 /2 7 /2 0 0 5 0 7 /2 7 /2 0 0 5 0 7 /2 7 /2 0 0 5 0 7 /2 7 /2 0 0 5 E05-0210 Interim R eport#13 Tennessee R iver Page 48 of 160 3M Environm ental Labora tory E05-0210 Interim R eport#13 Table 19. PFOA[1,2 13C] Surrogate Results Continued. 3 M Sam ple N um ber Sam ple Description (2)N o n -C o rre c te d (3)C o rrec te d (1)P F O A [1 ,2 13C ] S u rro g a te Surrogate C o n c e n tra tio n Percent Percent (ng/m L) Recovery Recovery E x tra c tio n D ate E 0 5 -0 2 1 0 -8 6 4 7 8 E 0 5 -0 2 1 0 -8 6 4 7 9 E 0 5 -0 2 1 0 -8 6 4 8 0 E 0 5 -0 2 1 0 -8 6 4 8 1 D X S -S W -T 0 2 0 0 3 -0 -0 5 0 7 2 1 D X S -S W -T 0 2 0 0 3 -0 -0 5 0 7 2 1 D u p D X S -S W -T 0 2 0 0 3 -0 -0 5 0 7 2 1 LS D X S -S W -T 0 2 0 0 3 -0 -0 5 0 7 2 1 H S 0 .1 4 9 0 .1 6 5 0 .1 3 6 0 .1 3 3 5 4 .4 6 0 .2 5 3 .5 5 2 .4 9 6 .9 107 9 5 .4 9 3 .3 0 7 /2 7 /2 0 0 5 0 7 /2 7 /2 0 0 5 0 7 /2 7 /2 0 0 5 0 7 /2 7 /2 0 0 5 E 0 5 -0 2 1 0 -8 6 4 8 2 E 0 5 -0 2 1 0 -8 6 4 8 3 E 0 5 -0 2 1 0 -8 6 4 8 4 E 0 5 -0 2 1 0 -8 6 4 8 5 D X S -S W -T 0 2 0 0 4 -0 -0 5 0 7 2 1 D X S -S W -T 0 2 0 0 4 -0 -0 5 0 7 2 1 D u p D X S -S W -T 0 2 0 0 4 -0 -0 5 0 7 2 1 LS D X S -S W -T 0 2 0 0 4 -0 -0 5 0 7 2 1 H S 0 .1 7 2 0 .1 5 4 0 .1 6 1 0 .1 3 1 6 2 .8 5 6 .2 5 8 .8 4 7 .8 112 100 105 8 5 .2 0 7 /2 7 /2 0 0 5 0 7 /2 7 /2 0 0 5 0 7 /2 7 /2 0 0 5 0 7 /2 7 /2 0 0 5 E 0 5 -0 2 1 0 -8 6 4 8 6 E 0 5 -0 2 1 0 -8 6 4 8 7 E 0 5 -0 2 1 0 -8 6 4 8 8 E 0 5 -0 2 1 0 -8 6 4 8 9 E 0 5 -0 2 1 0 -8 6 4 9 0 E 0 5 -0 2 1 0 -8 6 4 9 1 E 0 5 -0 2 1 0 -8 6 4 9 2 E 0 5 -0 2 1 0 -8 6 4 9 3 D X S -S W -T 0 2 0 0 5 -0 -0 5 0 7 2 1 D X S -S W -T 0 2 0 0 5 -0 -0 5 0 7 2 1 D u p D X S -S W -T 0 2 0 0 5 -0 -0 5 0 7 2 1 LS D X S -S W -T 0 2 0 0 5 -0 -0 5 0 7 2 1 H S D X S -S W -T 0 2 0 0 6 -0 -0 5 0 7 2 1 D X S -S W -T 0 2 0 0 6 -0 -0 5 0 7 2 1 D u p D X S -S W -T 0 2 0 0 6 -0 -0 5 0 7 2 1 LS D X S -S W -T 0 2 0 0 6 -0 -0 5 0 7 2 1 H S 0 .1 7 8 0 .1 7 0 0 .1 3 9 0 .1 7 7 0 .1 5 1 0 .1 5 6 0 .1 5 1 0 .1 4 3 7 0 .1 6 2 .0 5 4 .7 6 9 .7 5 5 .1 5 6 .9 5 9 .4 5 6 .3 125 110 9 7 .6 124 9 8 .2 101 106 100 0 7 /2 7 /2 0 0 5 0 7 /2 8 /2 0 0 5 c e n trifu g e d 0 7 /2 7 /2 0 0 5 0 7 /2 8 /2 0 0 5 c e n trifu g e d 0 7 /2 7 /2 0 0 5 0 7 /2 7 /2 0 0 5 0 7 /2 7 /2 0 0 5 0 7 /2 7 /2 0 0 5 E 0 5 -0 2 1 0 -8 6 4 9 4 E 0 5 -0 2 1 0 -8 6 4 9 5 E 0 5 -0 2 1 0 -8 6 4 9 6 E 0 5 -0 2 1 0 -8 6 4 9 7 D X S -S W -T 0 2 0 0 7 -0 -0 5 0 7 2 1 D X S -S W -T 0 2 0 0 7 -0 -0 5 0 7 2 1 D u p D X S -S W -T 0 2 0 0 7 -0 -0 5 0 7 2 1 LS D X S -S W -T 0 2 0 0 7 -0 -0 5 0 7 2 1 H S 0 .1 4 2 0 .1 5 4 0 .1 3 9 0 .1 4 6 5 1 .8 5 6 .2 5 4 .7 5 7 .5 9 2 .4 100 9 7 .6 102 0 7 /2 7 /2 0 0 5 0 7 /2 7 /2 0 0 5 0 7 /2 7 /2 0 0 5 0 7 /2 7 /2 0 0 5 E 0 5 -0 2 1 0 -8 6 4 9 8 E 0 5 -0 2 1 0 -8 6 4 9 9 E 0 5 -0 2 1 0 -8 6 5 0 0 E 0 5 -0 2 1 0 -8 6 5 0 1 D X S -S W -T 0 2 0 0 8 -0 -0 5 0 7 2 1 D X S -S W -T 0 2 0 0 8 -0 -0 5 0 7 2 1 D u p D X S -S W -T 0 2 0 0 8 -0 -0 5 0 7 2 1 LS D X S -S W -T 0 2 0 0 8 -0 -0 5 0 7 2 1 H S 0 .1 5 1 0 .1 6 1 0 .1 4 7 0 .1 2 8 5 5 .1 5 8 .8 5 7 .9 5 0 .4 9 8 .2 105 103 8 9 .8 0 7 /2 7 /2 0 0 5 0 7 /2 7 /2 0 0 5 0 7 /2 7 /2 0 0 5 0 7 /2 7 /2 0 0 5 E 0 5 -0 2 1 0 -8 6 5 0 2 E 0 5 -0 2 1 0 -8 6 5 0 3 E 0 5 -0 2 1 0 -8 6 5 0 4 E 0 5 -0 2 1 0 -8 6 5 0 5 D X S -S W -T 0 2 0 0 9 -0 -0 5 0 7 2 1 D X S -S W -T 0 2 0 0 9 -0 -0 5 0 7 2 1 D u p D X S -S W -T 0 2 0 0 9 -0 -0 5 0 7 2 1 LS D X S -S W -T 0 2 0 0 9 -0 -0 5 0 7 2 1 H S 0 .1 5 9 0 .1 5 1 0 .1 5 3 0 .1 3 8 5 8 .0 5 5 .1 6 0 .2 5 4 .3 103 9 8 .2 107 9 6 .8 0 7 /2 7 /2 0 0 5 0 7 /2 7 /2 0 0 5 0 7 /2 7 /2 0 0 5 0 7 /2 7 /2 0 0 5 E 0 5 -0 2 1 0 -8 6 5 0 6 E 0 5 -0 2 1 0 -8 6 5 0 7 E 0 5 -0 2 1 0 -8 6 5 0 8 E 0 5 -0 2 1 0 -8 6 5 0 9 D X S -S W -T 0 2 0 1 0 -0 -0 5 0 7 2 1 D X S -S W -T 0 2 0 1 0 -0 -0 5 0 7 2 1 D u p D X S -S W -T 0 2 0 1 0 -0 -0 5 0 7 2 1 L S D X S -S W -T 0 2 0 1 0 -0 -0 5 0 7 2 1 H S 0 .1 4 3 0 .1 4 3 0 .1 3 1 0 .0 8 6 5 2 .2 5 2 .2 5 1 .6 3 3 .8 9 3 .0 9 3 .0 9 1 .9 6 0 .2 0 7 /2 7 /2 0 0 5 0 7 /2 7 /2 0 0 5 0 7 /2 7 /2 0 0 5 0 7 /2 7 /2 0 0 5 E 0 5 -0 2 1 0 -8 6 5 1 0 E 0 5 -0 2 1 0 -8 6 5 1 1 E 0 5 -0 2 1 0 -8 6 5 1 2 E 0 5 -0 2 1 0 -8 6 5 1 3 D X S -S W -T 0 3 0 0 1 -0 -0 5 0 7 2 1 D X S -S W -T 0 3 0 0 1 -0 -0 5 0 7 2 1 D u p D X S -S W -T 0 3 0 0 1 -0 -0 5 0 7 2 1 LS D X S -S W -T 0 3 0 0 1 -0 -0 5 0 7 2 1 H S 0 .1 4 4 0 .1 4 6 0 .1 4 1 0 .1 2 7 5 2 .6 5 3 .3 5 5 .5 5 0 .0 9 3 .7 9 5 .0 9 9 .0 8 9 .1 0 7 /2 7 /2 0 0 5 0 7 /2 7 /2 0 0 5 0 7 /2 7 /2 0 0 5 0 7 /2 7 /2 0 0 5 E 0 5 -0 2 1 0 -8 6 5 1 4 E 0 5 -0 2 1 0 -8 6 5 1 5 E 0 5 -0 2 1 0 -8 6 5 1 6 E 0 5 -0 2 1 0 -8 6 5 1 7 D X S -S W -T 0 3 0 0 2 -0 -0 5 0 7 2 1 D X S -S W -T 0 3 0 0 2 -0 -0 5 0 7 2 1 D u p D X S -S W -T 0 3 0 0 2 -0 -0 5 0 7 2 1 LS D X S -S W -T 0 3 0 0 2 -0 -0 5 0 7 2 1 H S 0 .1 6 5 0 .1 4 2 0 .1 4 2 0 .1 2 6 6 0 .2 5 1 .8 5 5 .9 4 9 .6 107 9 2 .4 9 9 .6 8 8 .4 0 7 /2 7 /2 0 0 5 0 7 /2 7 /2 0 0 5 0 7 /2 7 /2 0 0 5 0 7 /2 7 /2 0 0 5 E05-0210 Interim R eport#13 Tennessee R iver Page 49 of 160 3M Environm ental Labora tory E05-0210 Interim R eport#13 Table 19. PFOA[1,2 13C] Surrogate Results Continued. 3 M Sam ple N um ber Sam ple Description (2)N o n -C o rre c te d (3)C o rrec te d (1)P F O A [1 ,2 13C ] S u rro g a te Surrogate C o n c e n tra tio n Percent Percent (ng/m L) Recovery Recovery E05-0210-86518 E05-0210-86519 E05-0210-86520 E05-0210-86521 DXS-SW-T03003-0-050721 DXS-SW-T03003-0-050721 Dup DXS-SW-T03003-0-050721 LS DXS-SW-T03003-0-050721 HS 0.153 0.136 0.142 0.124 55.8 49.6 55.9 48.8 99.5 88.5 99.6 87.0 E05-0210-86522 E05-0210-86523 E05-0210-86524 E05-0210-86525 DXS-SW-T03004-0-050722 DXS-SW-T03004-0-050722 Dup DXS-SW-T03004-0-050722 LS DXS-SW-T03004-0-050722 HS 0.154 0.142 0.146 0.135 56.2 51.8 57.5 53.1 100 92.4 102 94.7 E05-0210-86526 DXS-SW-T03005-0-050722 0.139 50.7 90.4 E05-0210-86527 DXS-SW-T03005-0-050722 Dup 0.048 17.7 31.5 E05-0210-86528 E05-0210-86529 DXS-SW-T03005-0-050722 LS DXS-SW-T03005-0-050722 HS 0.165 0.138 65.0 54.3 116 96.8 E05-0210-86530 E05-0210-86531 DXS-SW-T03006-0-050722 DXS-SW-T03006-0-050722 Dup 0.151 0.138 55.1 50.4 98.2 89.8 E05-0210-86532 DXS-SW-T03006-0-050722 LS 0.012 4.61 8.21 E05-0210-86533 DXS-SW-T03006-0-050722 HS 0.155 61.0 109 E05-0210-86534 E05-0210-86535 DXS-SW-T03007-0-050722 DXS-SW-T03007-0-050722 Dup 0.164 0.179 59.8 65.3 107 116 E05-0210-86536 E05-0210-86537 E05-0210-86538 E05-0210-86539 E05-0210-86540 E05-0210-86541 DXS-SW-T03007-0-050722 LS DXS-SW-T03007-0-050722 HS DXS-SW-T03008-0-050722 DXS-SW-T03008-0-050722 Dup DXS-SW-T03008-0-050722 LS DXS-SW-T03008-0-050722 HS 0.155 0.127 0.162 0.158 0.141 0.132 61.0 50.0 59.1 57.7 55.5 52.0 109 89.1 105 103 99.0 92.6 E05-0210-86542 E05-0210-86543 E05-0210-86544 E05-0210-86545 E05-0210-86546 E05-0210-86547 E05-0210-86548 E05-0210-86549 DXS-SW-T03009-0-050722 DXS-SW-T03009-0-050722 Dup DXS-SW-T03009-0-050722 LS DXS-SW-T03009-0-050722 HS DXS-SW-T03010-0-050722 DXS-SW-T03010-0-050722 Dup DXS-SW-T03010-0-050722 LS DXS-SW-T03010-0-050722 HS 0.161 0.144 0.147 0.132 0.148 0.140 0.145 0.127 58.8 52.6 57.9 52.0 54.0 51.1 57.1 50.0 105 93.7 103 92.6 96.3 91.1 102 89.1 E x tra c tio n D ate 07/27/2005 07/27/2005 07/27/2005 07/27/2005 07/27/2005 07/27/2005 07/27/2005 07/27/2005 07/27/2005 07/28/2005 c e n trifu g e d 07/27/2005 07/27/2005 07/27/2005 07/27/2005 07/28/2005 c e n trifu g e d 07/27/2005 07/27/2005 07/28/2005 c e n trifu g e d 07/27/2005 07/27/2005 07/27/2005 07/27/2005 07/27/2005 07/27/2005 07/27/2005 07/27/2005 07/27/2005 07/27/2005 07/27/2005 07/27/2005 07/27/2005 07/27/2005 E05-0210 Interim R eport#13 Tennessee R iver Page 50 of 160 3M Environm ental Labora tory E05-0210 Interim R eport#13 Table 19. PFOA[1,2 13C] Surrogate Results Continued. 3M Sample Number Sample Description (2>Non-Corrected (3>Corrected (1>PFOA [1,2 13C] Surrogate Surrogate Concentration Percent Percent (ng/mL> Recovery Recovery Extraction Date E05-0210-86550 E05-0210-86551 E05-0210-86552 DLS-SW-TRIP01 DLS-SW-TRIP01 LS DLS-SW-TRIP01 HS 0.139 0.138 0.117 50.7 50.4 46.1 90.4 89.8 82.1 07/27/2005 07/27/2005 07/27/2005 E05-0210-86553 E05-0210-86554 E05-0210-86555 D X S -S W -TR IP 02 DXS-SW -TRIP02 LS DXS-SW -TRIP02 HS 0.137 0.130 0.112 50.0 51.2 44.1 89.1 91.2 78.6 07/27/2005 07/27/2005 07/27/2005 E05-0210-86556 E05-0210-86557 E05-0210-86558 D X S -S W -TR IP 03 DXS-SW -TRIP03 LS DXS-SW -TRIP03 HS 0.144 0.128 0.129 52.6 50.4 50.8 93.7 89.8 90.5 07/27/2005 07/27/2005 07/27/2005 (1) All results and calculations in this table are presented with three significant figures. Sample concentrations, values may vary slightly from the raw data. (2) Surrogate concentration in sample/sample duplicates = 0.274 ng/mL. Surrogate concentration in low and high field matrix spikes = 0.254 ng/mL. (3) A correction factor of 56.1% was applied to all surrogate recoveries. The overall average raw surrogate recovery was used to determine the correction factor. _x x x_ N on - C o rre c te d S u rro g a te P e rc e n t R e c o v e ry C o r r e c t e d S u r r o g a t e P e r c e n t R e c o v e r y = --------------------------------------------------------------------------------------------- -- Statistical Methods and Calculations Statistical methods used to interpret sample results include averages and standard deviations. The Analyst software program calculated sample concentrations using resultant analyte peak areas and the established quadratic, 1/x weighted, calibration curve. None of the samples analyzed for this interim report required dilution. Sample calculations and equations used to report method accuracy and precision are described below. A c c u ra c y a n d P re c is io n E q u a tio n s Calculated Concentration * . LCS/Surrogate Percent Recovery --------------------------------------- * 100% Spike Concentration Sam ple Spike Recovery - (Spiked Sample Concentration - Average Concentration : Field Sample & Field Sample Dup.) * 1 0 0 % Spike Concentraton % nc,n RSD (,DRel1ative Standard ... . Deviation) = standard deviation of replicates nnn. ------------------------------------ ----------- * 100% replicate average % RPD (Relative Percent Difference) = -A--b--s--o--l-u--t-e---d-i-f-f-e--r-e--n--c--e--b--e--t-w---e--e-n---s--a--m--p--l-e--d--u- p--l-i-c-a--t-e--s- *100% average sample concentration E05-0210 Interim R eport#13 Tennessee R iver Page 51 of 160 3M Environm ental Labora tory E05-0210 Interim R eport#13 D e te rm in a tio n o f A n a ly tic a l U n c e rta in ty Both the accuracy (percent recovery) and precision (%RSD) of the lab control spikes are used to estimate the analytical uncertainty for a given analyte. For example, the overall accuracy and precision for PFBS based on LCS results was 107%3.49% (three significant figures). The measured precision (%RSD) is then used to determine the range of the accuracy. Example: 106.8*(0.0349) = 3.73 106.8 + 3.73 = 110.6; 107 - 3.73 = 103 Thus, LCS accuracy results range from 103% to 110.6%. The absolute difference of the low and high ends of this range, when compared 100%, are then calculated. 110.6%-100%= 10.6% 103%-100 = 3%. The most conservative (largest) absolute difference is then used as the analytical uncertainty for the given analyte. Therefore, the analytical uncertainty for PFBS is given as 10010.6% for this set of data. For PFHS, the overall project accuracy and precision of the the LCS results was 110%4.72%. This produced an estimated analytical uncertainty of 10015.2%. For PFOS, the overall project accuracy and precision was 108%%5.79% resulting in an estimated analytical accuracy of 10013.8%. Statement of Conclusion Sample results for PFBS, PFHS, and PFOS were presented in Table 1. Laboratory control spikes were used to determine the method accuracy and precision for each analyte. The accuracy and precision were then used to estimate the analytical uncertainty. Recoveries of field matrix spiked samples within 10030% demonstrated that the overall analytical method was appropriate for the matrix collected. Analytical accuracy for select PFOS results was adjusted to 10050% for samples whose associated field matrix spikes produced recoveries exceeding 10030%. List of Attachments Attachment A: Sample Chromatograms and Calibration Curves Attachm ent B: Extraction and Analytical Methods Attachment C: Protocol and Protocol Amendments E05-0210 Interim R eport#13 Tennessee R iver Page 52 of 160 3M Environmental Laboratory E05-0210 Interim Report#13 Signature Page We certify that this report is a true and complete representation of the data for this study: l/i/whA) I Michael A. Santojji Sponsor Representative ol Date E0S-0210 Interim Report#13 Tennessee River Page S3 o f 160 3M Environm ental Laboratory E05-0210 Interim R eport#13 At t a c h m e n t A: Sa m p l e Ch r o m a t o g r a m s a n d Ca l ib r a t io n Cu r v e s E05-0210 Interim R eport#13 Tennessee R iver Page 54 of 160 i :: - . V , v . ' ' ." - v;- .. . r f : : . e 3M Environm ental Laboratory E05-0210 Interim R eport#13 -- . ~ -- m- :: - : . . E05-0210 Interim R eport#13 Tennessee R iver Page 55 of 160 3M Environm ental Laboratory E05-0210 Interim R eport#13 E05-0210 Interim R eport#13 Tennessee R iver Page 56 of 160 3M Environm ental Laboratory E05-0210 Interim R eport#13 E05-0210 Interim R eport#13 Tennessee R iver Page 57 of 160 3M Environm ental Laboratory E05-0210 Interim R eport#13 E05-0210 Interim R eport#13 Tennessee R iver Page 58 of 160 .1. ' .... . 4 49 m 3M Environm ental Laboratory E05-0210 Interim R eport#13 Fie:"s050726swr | 2161 96 99 ' 10.0 10.1 10.2 ' 103 10.4 10.5 1015 10.7 106 109 11.0 11.1 11.2 11.3 11.4 11.5 116 E05-0210 Interim R eport#13 Tennessee R iver Page 59 of 160 3M Environm ental Laboratory E05-0210 Interim R eport#13 E05-0210 Interim R eport#13 Tennessee R iver Page 60 of 160 ... v : .. . r l ' - - - ", .. m 3M Environm ental Laboratory E05-0210 Interim R eport#13 '329 ' 13.0 ' 13.1 132 ' 13.3 ' 134 E05-0210 Interim R eport#13 Tennessee R iver Page 61 of 160 3M Environm ental Laboratory E05-0210 Interim R eport#13 SampeName:"s0S0726a034" SampeID:"M&-050726-7MethodBank-7' Fie:"sQS0726awir PeakName:"pO{ic)' Messies):"49S999.2amu.495.&i130.0amu.49S.9e00amu E05-0210 Interim R eport#13 Tennessee R iver Page 62 of 160 1 : : q '.: - v ...........64 - -;6 ;: \ :4 '5 f . .r .86 66 . d .: i' . - :8 . 3M Environm ental Laboratory E05-0210 Interim R eport#13 13---- '---- 133-- '---- 131-----'---- 133-- '---- 133-- '---- 131---- '---- 133-- '-----138---- '---- 133-- '-- 13-- ------15---- ----- 1440-- ------141---- '---- 113-- '---- 1443-- '---- 1444---- '---- 1445-- '---- 1446-- '---- 14T3---- '---- 1448-- '----- ;::- : ,.6 : ' :q ' . .... ,6 :::r ;:-,.: ; : . 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S T " ri r= :; q .;, :,~ es E05-0210 Interim R eport#13 Tennessee R iver Page 67 of 160 3M Environm ental Laboratory E05-0210 Interim R eport#13 E05-0210 Interim R eport#13 Tennessee R iver Page 68 of 160 I ....... V ' ." v : : . . ;: ; : / - . C -.; .m 1165 1.1.5 356. ' 29 3M Environm ental Laboratory E05-0210 Interim R eport#13 E05-0210 Interim R eport#13 Tennessee R iver Page 69 of 160 3M Environm ental Laboratory E05-0210 Interim R eport#13 E05-0210 Interim R eport#13 Tennessee R iver Page 70 of 160 3M Environm ental Laboratory E05-0210 Interim R eport#13 E05-0210 Interim R eport#13 Tennessee R iver Page 71 of 160 3M Environm ental Laboratory E05-0210 Interim R eport#13 E05-0210 Interim R eport#13 Tennessee R iver Page 72 of 160 3M Environm ental Laboratory E05-0210 Interim R eport#13 At t a c h m e n t B: Ex t r a c t io n a n d A n a l y t ic a l Me t h o d s E05-0210 Interim R eport#13 Tennessee R iver Page 73 of 160 3M Environm ental Laboratory E05-0210 Interim Report#13 3M Environmental Laboratory Method Determination of Perftuorinaied Acids, Alcohols, Amides, and Sulfonates In Water By Solid Phase Extraction and High Performance Liquid Chromatography/Mass Spectrometry Method Number: ETS-8-154.1 Adoption Date: 28 Apr 2000 Revision Date: 5 May, 2003 Effective Date: 5 May, 2003 Approved By: William K. Reagen Manager Date ETS-8-154.1 Page 1 of 17 Determination of Perfluorinated Compounds in Water Using SPE and LC/MS. E 05-0210 Interim Report#13 Tennessee River Page 74 of 160 3M Environm ental Laboratory E05-0210 Interim R eport#13 1 Scope and Application This method was validated for the collection, extraction, and analytical procedure for the determination of Perfluorooctane sulfonate (PFOS), Perfluorooctane sulfonylamide (FOSA), and Perfluorooctanoate (PFOA) in groundwater, surface water, and drinking water samples. This method may also be applied to the determination of other perfluorinated acids, alcohols, amides, and sulfonates in similar matrices, as long as the defined QC elements are satisfied and with the understanding that the method is not validated for compounds outside the scope of the original protocol This method is based in part on the report "Method of Analysis for the Determination of Perfluorooctane sulfonate (PFOS), Perfluorooctane sulfonylamide (PFOSA), and Perfluorooctanoate (POAA) in Water" (see Section 17), as developed and validated by Exygen Research (formerly Centre Analytical Laboratories, Inc.). 2 Method Summary Water samples are collected from a site of interest and shipped cold to an analytical facility. Perfluorinated acids, alcohols, amides, and sulfonates are extracted from 40mL water samples using C18 solid phase extraction (SPE) cartridges. The compounds are eluted from the C18 cartridge, using methanol. Separation, identification, and measurement are accomplished by high performance liquid chromatography/ tandem mass spectrometry (HPLC/MS/MS) analysis. High performance liquid chromatography/mass spectrometry (HPLC/MS) may be used if the defined QC elements are satisfied. The concentration of each identified component is measured by comparing the MS response of the quantitation ion produced by that compound to the MS response of the quantitation ion produced by the same compound in an extracted calibration standard (external standard). 3 Definitions 3.1 Analytical Sample A portion of an extracted Laboratory Sample prepared for analysis. 3.2 Calibration Standard A solution prepared from the Working Standard (WS) and extracted according to this method. The calibration standard solutions are used to calibrate the instrument response with respect to analyte concentration. 3.3 Duplicate Sample (DS) A DS is a separate aliquot of a sample, taken in the analytical laboratory that is extracted and analyzed separately with identical procedures. Analysis of DSs compared to that of the first aliquot give a measure of the precision associated with laboratory procedures, but not with sample collection, preservation, or storage procedures. 3.4 Field Blank Control Sample (FB) ASTM Type I water placed in a sample container in the laboratory and treated as a sample in all respects, including exposure to sampling site conditions, storage, preservation and all analytical procedures. The purpose of the FB is to determine if test substances or other interferences are present in the field environment. ETS-8-154.1 Page 2 of 17 Determination of Perfluorinated Compounds in Water Using SPE and LC/MS. E05-0210 Interim R eport#13 Tennessee R iver Page 75 of 160 3M Environm ental Laboratory E05-0210 Interim R eport#13 3.5 Field Duplicate (FD) A sample collected in duplicate at the same time as the sample and placed under identical circumstances and treated exactly the same throughout field and laboratory procedures. Analysis of FD compared to that of the first sample gives a measure of the precision associated with sample collection, preservation and storage, as well as with laboratory procedures. 3.6 Field Matrix Spike (FMS) A sample collected in duplicate to which known quantities of the target analytes are added in the field at the time of sample collection. Alternatively, the known quantity of target analytes may be added to the sample bottle in the laboratory before the bottles are sent to the field. A known, specific volume of sample must be added to sample container without rinsing. This may be accomplished by making a "fill to this level" line on the outside of the sample container. The FMS should be spiked at approximately 50-150% of the expected analyte concentration in the sample. If the expected range of analyte concentrations is unknown, a low and a high spike may be prepared to increase the likelihood that a spike at an appropriate range is made. The FMS is analyzed to ascertain if any matrix effects, interferences, or stability issues may complicate the interpretation of the sample analysis. 3.7 Field Spike Control Sample (FSCS) An aliquot of ASTM Type I water to which known quantities of the target analytes are added in the field at the time of sample collection (at an appropriate concentration to be determined by the project lead) or in the laboratory prior to the shipment of the collection bottles. The FSCS is extracted and analyzed exactly like a sample to determine whether a loss of analyte could be attributed to sample storage and/or shipment. A low and high FSCS may be appropriate when expected sample concentrations are not known. 3.8 Laboratory Control Sample (LCS) An aliquot of ASTM Type I water to which known quantities of the target analytes are added in the laboratory. Two levels are included, one at the LLOQ (approx. 25 pg/mL), the other at a concentration of approx. 100-250 pg/mL or another concentration to be determined by the project lead. The LCS is extracted and analyzed exactly like a laboratory sample to determine whether the methodology is in control, and whether the laboratory is capable of making accurate measurements at the required method detection limit and higher. 3.9 Laboratory Sample A portion of a sample received from the field for testing. 3.10 Limit of Detection (LOD) The LOD is the lowest concentration of an analyte that can be measured and reported with 99% confidence that the analyte concentration is greater than zero. If required, the LOD may be determined in several ways, including signal-to-noise ratio and statistical calculations. 3.11 Limit of Quantitation (LOQ) The LOQ for a dataset is the lowest concentration (LLOQ) or highest concentration (ULOQ) that can be reliably achieved within the specified limits of precision and accuracy during routine operating conditions. Note: For many analytes, the LLOQ analyte concentration is selected as the lowest non-zero standard in the calibration curve to simplify data reporting. Sample LLOQs are matrix-dependent. 3.12 Matrix Spike (MS) ETS-8-154.1 Page 3 of 17 Determination of Perfluorinated Compounds in Water Using SPE and LC/MS. E05-0210 Interim R eport#13 Tennessee R iver Page 76 of 160 3M Environm ental Laboratory E05-0210 Interim R eport#13 A matrix spike is an aliquot of a sample, to which known quantities of target analytes are added in the laboratory. The MS is extracted and analyzed exactly like a laboratory sample to determine whether the sample matrix contributes bias to the analytical results. The background concentrations of the analytes in the sample matrix must be determined in a separate aliquot and the measured values in the MS corrected for background concentrations. 3.13 Method Blank An aliquot of ASTM Type I water that is treated exactly like a laboratory sample including exposure to all glassware, equipment, solvents, and reagents that are used with other laboratory samples. The method blank is used to determine if test substances or other interferences are present in the laboratory environment, the reagents, or the apparatus. 3.14 Method Detection Limit (MDL) Determination A MDL is the statistically calculated minimum amount of an analyte that can be measured with 99% confidence that the reported value is greater than zero. One of several processes that may be used to establish a LOD value is found in 40 CFR Part 136 Appendix B. 3.15 Sample A sample is a small portion collected from a larger quantity of material intended to represent the original source material. 3.16 Spiking Stock Standard (SSS) A solution prepared from stock standards used to prepare the working standard. 3.17 Stock Standard (SS) A concentrated solution of a single analyte prepared in the laboratory with an assayed reference compound. 3.18 Working Standard (WS) A solution of several analytes prepared in the laboratory from SSs and diluted as needed to prepare calibration standards and other required analyte solutions. 4 Warnings and Cautions 4.1 Health and Safety The acute and chronic toxicity of the standards for this method have not been precisely determined; however, each should be treated as a potential health hazard. Unknown samples may contain high concentrations of volatile toxic compounds. Sample containers should be opened in a hood and handled with gloves to prevent exposure. The laboratory is responsible for maintaining a safe work environment and a current awareness of local regulations regarding the handling of the chemicals used in this method. A reference file of material safety data sheets (MSDS) should be available to all personnel involved in these analyses. 4.2 Cautions None 5 Interferences ETS-8-154.1 Page 4 of 17 Determination of Perfluorinated Compounds in Water Using SPE and LC/MS. E05-0210 Interim R eport#13 Tennessee R iver Page 77 of 160 3M Environm ental Laboratory E05-0210 Interim R eport#13 During extraction and analysis, major potential contaminant sources are reagents and solid phase extraction devices. All materials used in the analyses shall be demonstrated to be free from interferences under conditions of analysis by running method blanks. Parts and supplies that contain Teflon should be avoided due to the possibility of interference and/or contamination. These may include, but are not limited to: wash bottles, Teflon lined caps, autovial caps, HPLC parts, etc. The use of disposable micropipettes or pipettes to aliquot standard solutions is recommended to make calibration standards and matrix spikes. 6 Instrumentation, Supplies, and Materials Note: Brand names, suppliers, and part numbers are for illustrative purposes only. Equivalent performance may be achieved using apparatus and materials other than those specified here, but demonstration of equivalent performance that meets the requirements of this method is the responsibility of the laboratory performing the analysis. 6.1 Instrumentation Balance, analytical (display at least 0.0001g), Mettler HPLC/MS/MS or HPLC/MS system, as described in Section 10. 6.2 Supplies and Materials. Sample collection bottles-- LDPE (e.g., NalgeneTM) narrow-mouth bottles with screw cap. Note: Do not use Teflon bottles or Teflon lined caps. Coolers for sample shipment. Ice for sample shipment. Vacuum pump, Buchi. Visiprep vacuum manifold, Supelco. Sep Pak Vac 6cc (1g) tC18 cartridges (part # WAT 036795),Waters. 50mL disposable polypropylene centrifuge tubes, VWR. 15mL disposable polypropylene centrifuge tubes, VWR. Disposable micropipettes (50-100pL, 100-200pL), Drummond. Class A pipettes and volumetric flasks, various. Hypercarb drop-in guard column (4mm) (part # 844017-400), Keystone. Stand-alone drop-in guard cartridge holder, Keystone. 125mL LDPE narrow-mouth bottles, Nalgene. 2mL clear HPLC vial kit (cat # 5181-3400), Agilent/Hewlett Packard. Standard lab equipment (graduated cylinders, disposable tubes, etc.), various. 7 Reagents and Standards Note: Suppliers and catalog numbers are for illustrative purposes only. Equivalent performance may be achieved using chemicals obtained from other suppliers. Do not use a lesser grade of chemical than those listed. ETS-8-154.1 Page 5 of 17 Determination of Perfluorinated Compounds in Water Using SPE and LC/MS. E05-0210 Interim R eport#13 Tennessee R iver Page 78 of 160 3M Environm ental Laboratory E05-0210 Interim R eport#13 7.1 Chemicals Methanol (MeOH), HPLC grade, JT Baker, Catalog No. JT9093-2. Ammonium Acetate, Reagent grade, Sigma-Aldrich, Catalog No. A-7330. ASTM Type I Water, prepared in-house. Sodium Thiosulfate, Reagent grade, JT Baker. 7.2 Standards Potassium perfluorooctane sulfonate Perfluorooctane sulfonylamide Ammonium perfluorooctanoate Others as required. 7.3 Reagent Preparation 250mg/mL sodium thiosulfate solution -- Dissolve 25g of sodium thiosulfate in 100mL reagent water. 40% methanol wash solution - Measure 400mL methanol and adjust volume to 1.0L with reagent water. 100mM ammonium acetate solution (Analysis)--Weigh 7.71g of ammonium acetate and dissolve in 1.0L of reagent water. Dilute the 100mM solution by a factor of 50 to make the 2mM ammonium acetate solution used for mobile phase A. Note: Alternative volumes may be prepared as long as the ratios of the solvent to solute ratios are maintained. 7.4 Spiking Stock Standard (SSS) Preparation The following standard preparation procedure serves as an example and may be changed to suit the needs of a particular study. For example, pL volumes may be spiked into volumetric flasks when diluting stock solutions to appropriate levels. 100pg/mL each PFOS, PFOSA, and POAA SSSs--Weigh out 10mg of analytical standard (corrected for percent salt and purity--i.e., 10 mg C8F17S03K purity 90% = 8.35mg C8F17S03-) and dilute to 100mL with methanol in a 100mL volumetric flask. Transfer to a 125mL LDPE bottle or other suitable container. Prepare a separate solution for each analyte. Solutions may be stored in a refrigerator at 42C for a maximum period of 6 months from the date of preparation. 1pg/mL mixed SSS--Add 1.0mL each of the 100pg/mL SSSs (from 7.4.1) to a 100mL volumetric flask and bring up to volume with methanol. 0.1pg/mL mixed SSS--Add 10.0mL of the 1.0pg/mL-mixed solution (from 7.4.2) to a 100mL volumetric flask and bring up to volume with methanol. 0.01pg/mL mixed SSS--Add 10.0mL of the 0.1pg/mL-mixed solution (from 7.4.3) to a 100mL volumetric flask and bring up to volume with methanol. Storage Conditions--Store all SSSs in a refrigerator at 42C for a maximum period of 6 months from the date of preparation. 7.5 Calibration Standards The following standard preparation procedure serves as an example and may be changed to suit the needs of a particular study, provided the concentrations are calculated correctly. ETS-8-154.1 Page 6 of 17 Determination of Perfluorinated Compounds in Water Using SPE and LC/MS. E05-0210 Interim R eport#13 Tennessee R iver Page 79 of 160 3M Environm ental Laboratory E05-0210 Interim R eport#13 100pg/mL each PFOS, PFOSA, and POAA stock standard solutions--Weigh out 10mg of analytical standard (corrected for percent salt and purity) and dilute to 100mL with methanol in a 100mL volumetric flask. Transfer to a 125mL LDPE bottle or other suitable container. Prepare a separate solution for each analyte. Store solutions in a refrigerator at 42C for a maximum period of 6 months from the date of preparation. 1pg/mL Working Standard--Add 1.0mL each of the 100pg/mL SS solutions (from 7.5.1) to a 100mL volumetric flask and bring up to volume with methanol. 0.1pg/mL Working Standard --Add 10.0mL of the 1.0pg/mL mixed solution (from 7.5.2) to a 100mL volumetric flask and bring up to volume with methanol. 0.01pg/mL Working Standard --Add 10.0mL of the 0.1pg/mL mixed solution (from 7.5.3) to a 100mL volumetric flask and bring up to volume with methanol. Storage Conditions--Store all WSs in a refrigerator at 42C for a maximum period of 6 months from the date of preparation. Calibration Standard-- Prepare calibration solutions in ASTM Type I using the following table as a guideline: Final Calibration Concentration Volume of Standard Volume, mL , of WS, pg/mL WS, pL of ASTM Type I Water 0.0 0 40 0.010 100 40 0.010 200 40 0.010 400 40 0.10 100 40 0.10 200 40 0.10 300 0.10 400 40 40 1.0 100 1.0 400 40 40 1.0 1000 40 Final Concentration of Calibration Standard, pg/mL, in ASTM Type I Water 0 25 50 100 250 500 750 1000 2500 10000 25000 The standards are processed through the extraction procedure (Section 11), identical to the laboratory samples. The concentration of the calibration standard in the final extract is equal to 8X the initial concentration, due to the concentration of the standard during the extraction process. Storage Conditions-- Store all extracted calibration standards in 15mL polypropylene tubes at 42C, for a maximum period of two weeks from the date of preparation 8 Sample Collection and Handling Note: Sampling equipment, including automatic samplers, must be free of Teflon tubing, gaskets, and other parts that may leach interfering analytes into the water sample. Automatic samplers that composite samples over time should use refrigerated polypropylene sample containers if possible. Sample bottles should not be rinsed before sample collection. Labeling: Each sample bottle must display information regarding the collection of that sample, the individual collecting the sample, and any matrix spike that has been added to the sample. ETS-8-154.1 Page 7 of 17 Determination of Perfluorinated Compounds in Water Using SPE and LC/MS. E05-0210 Interim R eport#13 Tennessee R iver Page 80 of 160 3M Environm ental Laboratory E05-0210 Interim R eport#13 This includes the volume and concentration of any spiking solution added and the volume and identification of any preservatives added in the field. Spiking: The spiking scheme will be clearly outlined in the sampling plan, including whether the samples will be spiked in the field or in the laboratory prior to the shipment of the bottles to the site. If spiking is to be performed in the field, materials and specific instructions will be included in the sampling kit. Be sure to clearly label each bottle with spiking information if applicable. Tap Water: Open the tap and allow the system to flush until the water temperature (15o10C) has stabilized (usually about two minutes). Adjust the flow to about 500mL/min and collect samples from the flowing stream. Ground Water: Purge the well of standing water using a pump or a bailer. Collect the sample directly from the pump or from the bailer. Surface Water: When sampling from an open body of water, fill the sample container with water from a representative area. Sample Dechlorination: All samples should be iced or refrigerated at 42C and kept in the dark from the time of collection until extraction. Residual chlorine should be eliminated by adding 200pL of a 250mg/mL sodium thiosulfate solution to each tap-water sample and associated FB and FSCS (which may be placed in each bottle before leaving for the sampling site or done in the field.). Holding Time (HT): Results of the time/storage study of all target analytes showed that the three compounds are stable for 14 days in water samples when the samples are dechlorinated and stored as described in the previous section (see also references in section 17). Therefore, laboratory samples must be extracted within 14 days and the extracts analyzed within 30 days of sample collection. If the HT exceeds 14 days, great care is used when evaluating field spikes to avoid misrepresentation of the sample concentration. 8.1 Field Blanks Process a Field Blank Control Sample (FB) along with each sample set (samples collected from the same general sample site at approximately the same time). At the laboratory, prior to sample collection, fill a sample container with ASTM Type I water, seal, and ship the FB to the sampling site along with the empty sample containers. Return the FB to the laboratory with the filled sample bottles. When sodium thiosulfate is added to samples, use the same procedure to preserve the FB. 8.2 Field Duplicates Collect a Field Duplicate (FD) for every ten (10) samples collected or per each sampling set, if less than 10 samples are collected. Separate FDs must be collected for each type of water sample (ground, tap, etc.) collected. Collect the FD immediately after the sample. Preserve, store and ship FD using the same procedures as used for the samples. 8.3 Field Spike Control Sample (FSCS) A Field Spike Control Sample (FSCS) must be prepared for each sample shipment. If multiple coolers are used to ship a set of samples, each cooler must contain a FSCS. At the laboratory, fill a sample container with 100mL of ASTM Type I water. Seal and ship to the sampling site along with the empty sample containers and FBs. Samples may either be spiked in the field or in the laboratory prior to shipment. The method employed should be consistent throughout the study. If the samples are to be spiked in the field, be sure to send appropriate supplies and instructions for the field personnel to follow. ETS-8-154.1 Page 8 of 17 Determination of Perfluorinated Compounds in Water Using SPE and LC/MS. E05-0210 Interim R eport#13 Tennessee R iver Page 81 of 160 3M Environm ental Laboratory E05-0210 Interim R eport#13 Seal and gently invert the FSCS to mix. Store and ship the FSCS using the same procedures as used for the samples Provide information on sample collection, preservation, shipment and storage. List applicable holding times. Include sample stability and extract storage requirements. Reference the method used for sample preparation, if applicable. 8.4 Field Matrix Spike (FMS) A Field Matrix Spike (FMS) must be prepared for each sampling location. One unspiked sample from the same location must accompany the FMS to determine endogenous levels in the sample. The samples should be clearly identifiable as being from the same location. Samples may either be spiked in the field or in the laboratory prior to shipment. The method employed should be consistent throughout the study. If the samples are to be spiked in the field, be sure to send appropriate supplies and instructions for the field personnel to follow. 9 Quality Control and Data Quality Objectives Analytical results of the FB, FMS, FD, and FSCS should be evaluated at the conclusion of the study to help interpret the quality of sample data. Analytical results for these control/duplicate samples must be reported with the sample data. 9.1 Solvent Blanks Solvent blanks are analyzed with each sample set to determine contamination or carryover. Aliquots of methanol represent the solvent used for the standard curve and the sample extraction. Solvent blanks should have area counts that are less than 50% of the area count of the lowest calibration standard. Solvent blanks should be analyzed prior to and following each calibration curve, each set of system suitability samples, and after no more than 10 unknown sample extracts. If instrument carryover is a problem consecutive solvent blanks may be necessary. In this case the area counts of the solvent blanks should return to <50% of the lowest calibration standard prior to the injection of further standards or samples. 9.2 Method Blanks A method blank consists of an aliquot of ASTM Type I water, equal in volume to the samples, and extracted in the same manner as the samples. At least two method blanks should be prepared and analyzed each day that extractions are performed for a particular study or project. When analyzed the area counts of these samples must be less than 50% of the area count of the lowest calibration standard. 9.3 Sample Replicates All samples, including field spikes, trip blanks, etc., should be extracted at least in duplicate, and in triplicate if difficulties were encountered in the sampling and/or holding conditions of the samples. The relative percent difference (RPD) of duplicate samples or relative standard deviation (RSD) should be less than 15% for the precision of sample preparation and analysis to be considered in control. 9.4 Matrix Spike Matrix spikes are prepared for each sample type and analyzed to determine the matrix effect on the recovery efficiency. Matrix spike recoveries should fall within 25% of expected values. If the matrix spikes fail, evaluate the lab control spikes. If the LCS are within acceptance criteria there may be matrix issues in the samples. Discuss these in the final report. ETS-8-154.1 Page 9 of 17 Determination of Perfluorinated Compounds in Water Using SPE and LC/MS. E05-0210 Interim R eport#13 Tennessee R iver Page 82 of 160 3M Environm ental Laboratory E05-0210 Interim R eport#13 Matrix spike duplicates are prepared periodically to measure the precision associated with the analysis. Analyze a matrix spike and matrix spike duplicate (if prepared) in the same run as the original sample. Matrix spike and matrix spike duplicate concentrations should fall in the mid-range of the initial calibration curve or should be prepared at 1.5-5 times the endogenous concentration of the analyte. Spike concentrations should fall in the low-range of the initial calibration curve if extremely low-levels are expected. Generally two or more levels are prepared, one in the low range of the curve and one in the mid-range. This avoids the need to pre-screen unknown samples prior to preparation. 9.5 Laboratory Control Spike Lab control spikes are prepared for each study to ensure recovery of the target analytes. These should be prepared at a minimum of 2 levels and in duplicate or triplicate. Recovery of these samples should be within 25% of expected values, and the RPD (or RSD) be s 15%. If recoveries fall outside these limits the samples should be addressed in the final report. 10 Calibration and Standardization 10.1 Instrument Setup Note: In this example, a MicroMass UltimaTM Liquid Chromatography Tandem Mass Spectrometer (LC/MS/MS) is used. Other brands of LC/MS/MSs as well as single quadrupole mass spectrometers (LC/MS) may be used as long as the method criteria are met. Brand names, suppliers, part numbers, and models are for illustrative purposes only. Equivalent performance may be achieved using apparatus and materials other than those specified here, but demonstration of equivalent performance that meets the requirements of this method is the responsibility of the laboratory. The operator must optimize and document the equipment and settings used. Establish the LC/MS/MS system and operating conditions equivalent to the following: Mass Spec: Micromass Ultima (Micromass) Interface: Electrospray (Micromass) Mode: Electrospray Negative, Multiple Response Monitoring (MRM) Harvard infusion pump (Harvard Instruments), for tuning Computer: COMPAQ Professional Workstation AP200 Software: Windows NT, MassLynx 3.3 HPLC: Hewlett Packard (HP) Series 1100 HP Quaternary Pump HP Vacuum Degasser HP Autosampler HP Column Oven Note: A 4 x 10mm Hypercarb drop-in guard cartridge (Keystone, part # 844017-400) may be attached on-line after the purge valve and before the sample injector port to trap any residue contaminants that may be in the mobile phase and/or HPLC system. HPLC Column: Genesis C8 (Jones Chromatography), 2.1mm x 50mm, 4pm ETS-8-154.1 Page 10 of 17 Determination of Perfluorinated Compounds in Water Using SPE and LC/MS. E05-0210 Interim R eport#13 Tennessee R iver Page 83 of 160 3M Environm ental Laboratory E05-0210 Interim R eport#13 Column Temperature: 35C Injection Volume: 15pL Mobile Phase (A): 2mM Ammonium Acetate in ASTM Type I water (See 7.3.1) Mobile Phase (B): Methanol Time, Percent Mobile Percent Mobile Flow Rate, min Phase A Phase B mL/min 0.0 60 40 0.3 0.4 60 40 0.3 1.0 10 90 0.3 7.0 10 90 0.3 7.5 0 100 0.3 9.0 0 100 0.4 9.5 60 40 0.4 13.5 60 40 0.4 14.0 60 40 0.3 Note: Other HPLC gradients may be used as long as the method criteria are met. It may be necessary to adjust the HPLC gradient in order to optimize instrument performance. Columns with different dimensions (e.g. 2.1mm x 30mm) and columns from different manufacturers (Keystone Betasil C18 etc.) may be used. Ions Monitored: Analyte Primary Ion Product Ion Approximate Retention Time (minutes) PFOA PFOS FOSA 413 499 498 169 99 78 5.0 5.2 5.8 Other product ions may be chosen at the discretion of the analyst, although m/z 99 is suggested for PFOS. Use of the suggested primary ion is recommended. Retention times may vary slightly, on a day-to-day basis, depending on the batch of mobile phase etc. Drift in retention times is acceptable within an analytical run, as long as the drift continues through the entire analysis and the standards are interspersed throughout the analytical run. 10.2 Tune File Parameters The following values are provided as an example. Actual values may vary from instrument to instrument. Also, these values may be changed from time to time in order to optimize for greatest sensitivity. Analyte PFOA PFOS FOSA Dwell, sec 0.2-0.4 0.2-0.4 0.2-0.4 Collision Energy, eV 10-25 30-60 20-50 Cone, V 20-30 50-80 30-60 ETS-8-154.1 Page 11 of 17 Determination of Perfluorinated Compounds in Water Using SPE and LC/MS. E05-0210 Interim R eport#13 Tennessee R iver Page 84 of 160 3M Environm ental Laboratory E05-0210 Interim R eport#13 Source Capillary Hexapole 1 Aperture 1 Hexapole 2 Source Block Temp. Desolvation Temp. Analyzer LM Res 1 HM Res 1 lEnergy 1 Entrance Exit LM Res 2 HM Res 2 lEnergy 2 Multiplier Gas Flows Cone Gas Desolvation Pressures Gas Cell Set 2.6-3.5kV 0.5V 0.2V 0.8V 100-150C 250-400C Set 12.5-15.0V 12.5-15.0V 0.7V -2V 1V 11.0V 11.0V 1.0V 650V Set 150L/hr 700L/hr Set 3.0e-3mbar ETS-8-154.1 Page 12 of 17 Determination of Perfluorinated Compounds in Water Using SPE and LC/MS. E05-0210 Interim R eport#13 Tennessee R iver Page 85 of 160 3M Environm ental Laboratory E05-0210 Interim R eport#13 10.3 Calibration Curve Analyze the standard curves prior to each set of samples. The validated method specifies that the standard curve should be plotted using a linear fit, weighted 1/x or unweighted. However, the standard curve may also be plotted by quadratic fit (y = ax'2 + bx + c), weighted 1/x or unweighted, using suitable software. The calibration curves may include but should not be forced through zero. The mathematical method used to calculate the calibration curve should be applied consistently throughout a study. Any change should be thoroughly documented in the raw data. If the calibration curve does not meet acceptance criteria perform routine maintenance or prepare a new standard curve (if necessary) and reanalyze. For purposes of accuracy when quantitating low levels of analyte, it may be necessary to use the low end of the calibration curve rather than the full range. For example, when attempting to quantitate approximately 50 pg/mL of analyte, generate a calibration curve consisting of the standards from 25 pg/mL to 1000 pg/mL rather than the full range of the curve (25 pg/mL to 25000 pg/mL). This will reduce inaccuracy attributed to linear regression weighting of high concentration standards. High and/or low points may be excluded from the calibration curves to provide a better fit over the linear range appropriate to the data or because they did not meet the pre-determined acceptance criteria. Low-level curve points should also be excluded if their area counts are not at least twice that of the method and/or solvent blanks. Any curve point may be rejected due to a bad injection or failing to meet accuracy requirements of 25% (and 30% for the LLOQ). Justification for exclusion of calibration curve points will be noted in the raw data. A minimum of 6 points will be used to construct the calibration curve. 10.4 Continuing Calibration Verification (CCV) Continuing calibration verifications (CCV) are analyzed to verify the accuracy of the calibration curve. Analyze a mid-range calibration standard, one of the same standards used to construct the calibration curve, at a minimum after every tenth sample, not including solvent blanks, with a minimum of one per sample set. Calibration verification injections must be within 25% to be considered acceptable. The calibration curve and the last passing CCV will then bracket acceptable samples. Multiple CCV levels may be used. 10.5 System Suitability A minimum of three system suitability samples will be injected at the beginning and end of each analytical run. Typically these samples are run prior to the calibration curve. The system suitability injections must have area counts with an RSD of s5% and a retention time RSD of s2% when evaluated independently. 11 Procedures 11.1 Extraction Scheme Allow samples to equilibrate to room temperature. Thoroughly mix samples by gently inverting the sample bottle. Measure 40mL of sample into 50mL polypropylene centrifuge tubes (Spike the Matrix spikes as required*, replace lid and mix well). Note: * Samples may need to be prescreened to determine an appropriate matrix spike level (typically 50-150% of sample concentration). Alternatively the samples could be spiked at more than one level, allowing for the inappropriate spike level to be eliminated. ETS-8-154.1 Page 13 of 17 Determination of Perfluorinated Compounds in Water Using SPE and LC/MS. E05-0210 Interim R eport#13 Tennessee R iver Page 86 of 160 3M Environm ental Laboratory E05-0210 Interim R eport#13 Condition the C18 SPE cartridges (1g, 6mL) by passing approximately 10mL methanol followed by approximately 50mL ASTM Type I water (flow rate approximately 2 drop/sec). Do not let column run dry. Note: For the following steps, maintain a ~1drop/sec flow rate. Do not allow the column to run dry at any time. Load the analytical sample onto the C18 SPE cartridge. Discard eluate. Ten mL of the 40% methanol in water wash mixture is passed through the C18 SPE cartridge to rinse away potential interferences and then discarded. This step must be omitted if perfluorinated compounds with chain lengths less than C8 are targeted since these will be lost during this wash step. Elute with exactly 5mL of 100% methanol. Collect eluate into graduated 15mL polypropylene centrifuge tubes. This is the target elution fraction (final volume approximately 4.5 mL as not all of the solvent will leave the SPE column. This will not affect the calculations in any way since the curve is also extracted). Analyze a portion of the target elution fraction eluent using negative electrospray HPLC/MS/MS or HPLC/MS. Note: Samples are concentrated by a factor of eight during the extraction; Initial Vol = 40mL ^ Final Vol. = 5mL. Samples are stable at room temperature for at least 24 hours. Analytical samples may be stored in a refrigerator at 42C until analysis. Standardization of C18 SPE columns-- If poor recoveries are observed, it may be necessary to standardize the C18 SPE columns in the following manner before analyzing samples. Use a standard with an analyte concentration between 1000 and 4000 pg/mL. Repeat the extraction scheme from the beginning up through the eluting with ~5mL 100% methanol. After the eluting with ~5mL 100% methanol step, collect an additional post-elution fraction by eluting with an additional 5mL of 100% methanol. Analyze both fractions by HPLC/MS/MS or HPLC/MS. If the target fraction contains a minimum of 85% of the respective analytes, it may be considered acceptable. If the wash contains significant standard (>15%), either the wash volume or percentage of MeOH should be decreased. If the post-elution fraction contains significant standard (>15%), the target elution volume should be increased. 11.2 Sample Analysis Set up analysis sample queue. Inject the same volume (between 5-25pL) of each standard, analytical sample and blank into the instrument. All samples with a concentration > ULOQ must be diluted and reanalyzed. If dilution of the final extract fails to produce acceptable results (e.g. poor MS recoveries) dilute the original sample and re-extract. 12 Data Analysis and Calculations Calculate the analytical sample (extract) concentration from the standard curve using the following equation: ETS-8-154.1 Page 14 of 17 Determination of Perfluorinated Compounds in Water Using SPE and LC/MS. E05-0210 Interim R eport#13 Tennessee R iver Page 87 of 160 3M Environm ental Laboratory E05-0210 Interim R eport#13 Extract Concentration, pg/mL = (Peak area - intercept) (slope) Calculate the percent recovery of the FSCS using the following equation: FSCS % rec. = (FSCS conc., pg/mL) x 100 (Conc. added, pg/mL) Calculate the percent recovery of the MSs using the following equation: MS % rec. = (MS conc., pg/mL - Sample conc., pg/mL) x 100 (Conc. added, pg/mL) 13 Method Performance Note: Any method performance parameters that are not achieved must be considered in the evaluation of the data. Nonconformance to any specified parameters must be described and discussed in any reporting of the data. If criteria listed in this method performance section are not met, maintenance may be performed on the system and samples reanalyzed, or other actions taken as determined by the analyst. Document all actions in the raw data. If data are to be reported when performance criteria have not been met, the data must be footnoted on tables and discussed in the text of the report. 13.1 System Suitability A minimum of three system suitability samples will be injected at the beginning and end of each analytical run. Typically these samples are run prior to the calibration curve. The system suitability injections must have area counts with an RSD of s5% and a retention time RSD of s2% when evaluated independently. 13.2 Quantitation Calibration Curve: The coefficient of determination (r2) value for the calibration curve must be greater than or equal to 0.990. Each point in the curve must be within 25% of the theoretical concentration with the exception of the LLOQ, which may be within 30%. Demonstration of Specificity: Specificity is demonstrated by chromatographic retention time (within 3% of standard) and the mass spectral response of unique ions. 13.3 Sensitivity Solvent Blanks and Method Blanks: Solvent and method blank area counts must be < 50% that of the lowest standard used in the calibration curve. Limits of Quantitation (LOQ): The lower LOQ (LLOQ) is the lowest non-zero active standard in the calibration curve; the peak area of the LLOQ must be at least 2X that of the extraction blank. By definition, the measured value of the LLOQ must be within 30% of the theoretical value. ETS-8-154.1 Page 15 of 17 Determination of Perfluorinated Compounds in Water Using SPE and LC/MS. E05-0210 Interim R eport#13 Tennessee R iver Page 88 of 160 3M Environm ental Laboratory E05-0210 Interim R eport#13 13.4 Accuracy CCV Performance: Calibration verification injections must be within 25% to be considered acceptable. The calibration curve and the last passing CCV will then bracket acceptable samples. Multiple CCV levels may be used. Matrix Spikes: Matrix spike percent recoveries must be within 25% of the spiked concentration. If matrix effects are suspected, evaluate the LCS results to determine if a matrix effects are present and if the method is in control based on compliant LCS results. Discuss all results in the analytical report. 13.5 Precision Reproducibility: Reproducibility of the method is defined by the results of duplicate or triplicate analysis of samples. A RPD or RSD of s 15% will be considered acceptable. System Suitability: The system suitability injections must have area counts with an RSD of s5% and a retention time RSD of s2% when evaluated independently. 14 Pollution Prevention and Waste Management Sample extract waste and flammable solvent is discarded in high BTU containers, and glass pipette waste is discarded in broken glass containers located in the laboratory. 15 Records Each data package generated for a study must have the following information included: study or project number, acquisition method, integration method, sample name, extraction date, dilution factor (if applicable), and analyst. Print the tune page, sample list, and acquisition method to include in the appropriate study folder. Copy these pages and tape into the instrument run log. Plot the calibration curves as described in this method, then print these graphs and store in the study folder. Print data integration summary, integration method, and chromatograms and store in the study folder. Summarize data using suitable software and store in the study folder. 16 Attachments None. 17 References "Method of Analysis for the Determination of Perfluorooctane sulfonate (PFOS), Perfluorooctane sulfonylamide (PFOSA), and Perfluorooctanoate (POAA) in Water", E. Wickremesinhe and J. Flaherty, Study Number 023-002, Centre Analytical Laboratories, Inc., State College, Pennsylvania, January 2000. Validation report for the "Method of Analysis for the Determination of Perfluorooctane sulfonate (PFOS), Perfluorooctane sulfonylamide (PFOSA), and Perfluorooctanoate (POAA) in Water", E. Wickremesinhe and J. Flaherty, Study Number 023-002, Centre Analytical Laboratories, Inc., State College, Pennsylvania. ETS-8-154.1 Page 16 of 17 Determination of Perfluorinated Compounds in Water Using SPE and LC/MS. E05-0210 Interim R eport#13 Tennessee R iver Page 89 of 160 3M Environm ental Laboratory E05-0210 Interim R eport#13 18 Affected Documents None. 19 Revisions Revision Number 1 Revision Number U pdated to the ne w form at. C hanged Title. S ection 1: S tates the validation o f 3 analytes, re m o ve s re fe re n ce to EPA docum ent that's no longer applicable. Section 2: Provided for the extraction o f m ore than the 3 validated analytes, allows the use o f a LC/M S system, not only the LS/M S/M S previously m e n tio n e d . Section 3: R evised definitions for field m atrix spike, field control spike, LLOQ, m ethod blank, and MDL. Section 5: R ew orded the interferences, added recom m endation to use disposable pipettes. Section 6: R ecategorized and pared down. Section 7: C hanged storage tim e to 6 m onths. A d ded m ore calibration points to the table. Section 8:Added statem ent addressing labeling requirem ents and spiking procedures. E xpanded section 8.8. Section 9: N ew Section Section 10: C hanged som e o f the param eters in the tables. A llow ed for use o f different instrum entation. A dded inform ation from section 12 o f previous version, extensively revised. S ection 11 (sectio n 9 in p re vio u s version): C larifica tio n o f w ash step, sta te d exact volum e o f eluate is 5 mL, revised standardization process, rem oved requirem ent to use LC/M S/M S. Section 12 (section 13 in previous version: no changes Section 13 (section 14 in previous version): E xtensively rewritten. Section 14 (section 15 in previous version): no changes Section 15 (section 16 in previous version): M inor changes to recording requirem ents. Section 16 (section 17 in previous version): R em oved attachment. Section 17 (section 18 in previous version): R em oved reference to EPA docum ent that no longer applied to this SOP. Section 18: N ew section. Revision Date 7 ETS-8-154.1 Page 17 of 17 Determination of Perfluorinated Compounds in Water Using SPE and LC/MS. E05-0210 Interim R eport#13 Tennessee R iver Page 90 of 160 3M Environm ental Laboratory E05-0210 Interim R eport#13 At t a c h m e n t C: Pr o t o c o l a n d Pr o t o c o l A m e n d m e n t s E05-0210 Interim R eport#13 Tennessee R iver Page 91 of 160 3M Environm ental Laboratory E05-0210 Interim Report#13 Exygen Protocol Number: P0001131 STUDY PROTOCOL Study Title: Analysis of Perfluorobutanesulfonate (PFBS), Perfluorohexanesulfonate (PFHS), and Perfluorooctanesulfonate (PFOS) in Water, Soil, Sediment, Fish, Clams, Vegetation, Small Mammal Liver and Small Mammal Serum Using LC/MS/MS for the 3M Decatur Monitoring Program Exygen Protocol Number: P0001131 Performing Laboratory: Exygen Research 3058 Research Drive State College, PA 16801 Phone: (814) 272-1039 Sponsor Representative: Michael A. Santoro Director of Regulatory Affairs 3M Building 0236-01-B-10 St. Paul, MN 55144 Phone: (651) 733-6374 E05-0210 Interim Report#13 Tennessee River Page / oj 65 Page 92 of 160 3M Environm ental Laboratory E05-0210 Interim Report#13 Exygen Protocol Number: P0001131 D IS TR IB U TIO N : 1) Jaisimha Kesari, Study Director, Weston Solutions 2) John M. Flaherty, Principal Investigator, Exygen Research 3) Michael A. Santoro, Sponsor Representative, 3M Company 4) Exygen Research Quality Assurance Unit E05-0210 Interim Report#13 Tennessee River Page 2 o f 65 Page 93 of 160 3M Environm ental Laboratory E05-0210 Interim Report#13 Exygen Protocol Number: P0001131 PROTOCOL APPROVAL Study Title: Analysis of Perfluorobutanesulfonate (PFBS), Perfluorohexanesulfonate (PFHS), and Perfluorooctanesulfonate (PFOS) in Water, Soil, Sediment, Fish, Clams, Vegetation, Small Mammal Livers and Small Mammal Serum Using LC/MS/MS for the 3M Decatur Monitoring Program Exygen Protocol Number: P0001131 _____________ _______________ Michael A. Santoro, Sponsor Representative 3M Compaify bhn M. Flaherty, Principal Investigator Exygen Research Richard A. Grafzzini, Pi^sident, Facility Management Exygen Research Lydua Shaffer, Technip^i^ead, Quality Assurance Unit Exygen Research E05-0210 Interim Report#13 Tennessee River uno )at Date - o c r - ' z f Date //zq /os Date Page 3 o f 65 Page 94 of 160 3M Environm ental Laboratory E05-0210 Interim Report#13 Exygen Protocol Number: P0001131 TABLE OF CONTENTS TITLE PA G E ........................................................................................................... DISTRIBUTION..................................................................................................... PROTOCOL APPROVAL.................................................................................... TABLE OF CONTENTS...................................................................................... INTRODUCTION................................................................................................... TEST M ATERIALS.............................................................................................. O B J E C T I V E ............................................................................................................ TESTING FACILITY............................................................................................ STUDY DIRECTOR.............................................................................................. SPONSOR REPRESENTATIVE........................................................................ PRINCIPAL INVESTIGATOR........................................................................... PROPOSED EXPERIMENTAL START AND TERMINATION DATES IDENTIFICATION AND JUSTIFICATION OF THE TEST SYSTEM .... SAMPLE PROCUREMENT, RECEIPT AND RETENTION..................... SAMPLE IDENTIFICATION............................................................................. ANALYTICAL PROCEDURE SUM M ARY................................................... VERIFICATION OF ANALYTICAL PROCEDURE.................................... METHOD FOR CONTROL OF B IA S .............................................................. STATISTICAL M ETHODS................................................................................ GLP STATEM ENT............................................................................................... R E P O R T .................................................................................................................. SAFETY AND HEALTH..................................................................................... AMENDMENTS TO PROTOCOL.................................................................... DATA RECORD KEEPING............................................................................... QUALITY A SSU R A N C E .................................................................................... RETENTION OF DATA AND ARCHIVING................................................. APPENDIX I, ANALYTICAL METHODS..................................................... 1 2 -> 4 5 6 6 7 7 7 7 8 8 9 9 9 11 11 11 11 12 13 13 14 14 15 E05-0210 Interim Report#13 Tennessee River Page 4 o f 65 Page 95 of 160 3M Environm ental Laboratory E05-0210 Interim Report#13 Exygen Protocol Number: P0001131 IN TR O D U C TIO N The purpose of this study is to perform analysis for perfluorobutanesulfonate (PFBS), perfluorohexanesulfonate (PFHS) and perfluorooctanesulfonate (PFOS) in water, soil, sediment, fish, clams, vegetation, small mammal livers and small mammal serum using LC/MS/MS for the 3M Decatur Monitoring Program. The study will be audited for compliance with EPA TSCA Good Laboratory Practice Standards 40 CFR 792 by the Quality Assurance Unit of Exygen Research. TEST M A TE R IA LS The test materials are perfluorobutanesulfonate (PFBS), perfluorohexanesulfonate (PFHS) and perfluorooctanesulfonate (PFOS) and are all supplied by 3M. PFBS Chemical Name: Perfluorobutanesulfonate Molecular Weight: 338 supplied as the potassium salt (C4FqS 0 3'K+) Lot Number: 101 Purity: 96.7% Transitions Monitored: 299 -> 99 Structure: PFHS Chemical Name: Perfluorohexanesulfonate Molecular Weight: 438 supplied as the potassium salt (C6Fi3S0 3 'K+) Lot Number: SE036 Purity: 98.6% Transitions Monitored: 399 - 80 Structure: E05-0210 Interim Report#13 Tennessee River Page 5 o f 65 Page 96 of 160 3M Environm ental Laboratory E05-0210 Interim Report#13 Exygen Protocol Number: P0001 PFOS Chemical Name: Perfluorooctanesulfonate Molecular Weight: 538 supplied as the potassium salt (C8Fi7S 0 3'K+) Lot Number: 217 Purity: 86.9% Transitions Monitored: 499 -- 99 Structure: O B JEC TIVE The purpose of this study is to perform analysis for perfluorobutanesulfonate (PFBS), perfluorohexanesulfonate (PFHS) and perfluorooctanesulfonate (PFOS) in water, soil, sediment, fish, clams, vegetation, small mammal livers and small mammal serum for the 3M Decatur Monitoring Program using the current versions of the following Exygen analytical methods: V0001780: "Method of Analysis for the Determination of Perfluorooctanoic Acid (PFOA) in Water by LC/MS/MS" V0001781: "Method of Analysis for the Determination of Perfluorooctanoic Acid (PFOA) in Soil by LC/MS/MS" V0001782: "Method of Analysis for the Determination of Perfluorooctanoic Acid (PFOA) in Sediment by LC/MS/MS" V0001783: "Method of Analysis for the Determination of Perfluorooctanoic Acid (PFOA) in Fish and Clams by LC/MS/MS" V0001784: "Method of Analysis for the Determination of Perfluorooctanoic Acid (PFOA) in Vegetation by LC/MS/MS" V0001785: "Method of Analysis for the Determination of Perfluorooctanoic Acid (PFOA) in Small Mammal Liver by LC/MS/MS" V0001786: "Method of Analysis for the Determination of Perfluorooctanoic Acid (PFOA) in Small Mammal Serum by LC/MS/MS" TE S TIN G F A C IL IT Y Exygen Research 3058 Research Drive State College, PA 16801 Phone: (814) 272-1039 Page 6 o f 65 E05-0210 Interim Report#13 Tennessee River Page 97 of 160 3M Environm ental Laboratory E05-0210 Interim Report#13 Exygen Protocol Number: P0001131 STUDY D IR E C TO R Jaisimha Kesari P.E., DEE Weston Solutions, Inc. 1400 Weston Way West Chester, PA 19380 Phone: (610) 701-3761 Fax: (610) 701-7401 j .kesari@westonsolutions.com SPONSOR R E PR ESEN TA TIVE Michael A. Santoro 3M Company Director of Regulatory Affairs 3M Building 0236-01-B-10 St. Paul, MN 55144 Phone: (651) 733-6374 P R IN C IPA L IN V E S T IG A TO R John M. Flaherty Exygen Research 3058 Research Drive State College, PA 16801 Phone: (814) 272-1039 j ohn. flaherty@exygen. com PROPOSED E X P E R IM E N TA L START AND T E R M IN A T IO N DATES It is proposed that the analytical portion of this study be conducted from October 01, 2004 to December 31, 2005. The actual experimental start and termination dates will be included in the final report. E05-0210 Interim Report#13 Tennessee River Page 7 o f 65 Page 98 of 160 3M Environm ental Laboratory E05-0210 Interim Report#13 Exygen Protocol Number: P 0001131 ID E N T IF IC A T IO N AND JU S TIFIC A TIO N OF TH E TEST SYSTEM The following are the test systems for this study: Water (groundwater and surface water) Soil Sediment Fish Clams Vegetation Small Mammal Liver Small Mammal Serum The samples will be collected by Weston Solutions. The control samples will be purchased and prepared by the testing facility. Purchase and processing details for the control samples will be included in the final report associated with this study. The test systems were chosen to access the environmental impact of PFBS, PFHS and PFOS in the Decatur, Alabama area. SAM PLE PR O C UR EM EN T, R E C E IP T AND R E TE N TIO N Water, soil, sediment, fish, clam, vegetation, small mammal liver and small mammal serum samples will be received at Exygen directly from Weston Solutions. The details of sample procurement for this study are outlined in the 3M work plan entitled "Phase 2 Work Plan for Sampling Environmental Media." The number and types of samples collected will vary depending availability in the field. The total number of samples received and analyzed for each matrix will be documented in the final report associated with this study. Water, soil, and sediment samples will be used as received without further processing at Exygen. These samples will be stored refrigerated at 2C-8C. Fish, clam, vegetation and small mammal liver samples will be processed according to the appropriate analytical method (see Appendix I). These samples will be stored frozen at < -10C. Small mammal whole blood samples will be centrifuged in the field at the time of collection and the serum fraction will be used for the study. Small mammal serum will be stored frozen at < -10C. The receipt and processing of the samples will be documented in the final report and raw data associated with the study. E05-0210 Interim Report#13 Tennessee River Page 8 o f 65 Page 99 of 160 3M Environm ental Laboratory E05-0210 Interim Report#13 Exygen Protocol Number: P0001131 SAMPLE ID E N T IF IC A T IO N Prior to analysis, each sample will be assigned a laboratory sample reference number. The reference number will be unique and will distinguish each laboratory sample that is processed throughout the analytical procedure. Chromatographic data will be identified by the laboratory sample reference number. Sample storage conditions and locations will be documented throughout the study. A N A LY TIC A L PROCEDURE SU M M ARY References: V0001780: "Method of Analysis for the Determination of Perfluorooctanoic Acid (PFOA) in Water by LC/MS/MS" V0001781: "Method of Analysis for the Determination of Perfluorooctanoic Acid (PFOA) in Soil by LC/MS/MS" V0001782: "Method of Analysis for the Determination of Perfluorooctanoic Acid (PFOA) in Sediment by LC/MS/MS" V0001783: "Method of Analysis for the Determination of Perfluorooctanoic Acid (PFOA) in Fish and Clams by LC/MS/MS" V0001784: "Method of Analysis for the Determination of Perfluorooctanoic Acid (PFOA) in Vegetation by LC/MS/MS" V0001785: "Method of Analysis for the Determination of Perfluorooctanoic Acid (PFOA) in Small Mammal Liver by LC/MS/MS" V0001786: "Method of Analysis for the Determination of Perfluorooctanoic Acid (PFOA) in Small Mammal Serum by LC/MS/MS" The above methods use analytical conditions capable of separating the isomers of PFBS, PFHS and PFOS. The final report will include the isomers summed into total PFBS, total PFHS, and total PFOS found. V E R IF IC A T IO N OF A N A LY TIC A L PROCEDURE A laboratory control sample will be used for the preparation of fortified control samples. The test substance will be made into solutions as per the method, and added to the matrices via a micropipette. For water sampling, Exygen will supply one bottle per sample collected. The bottles will be 500 mL precleaned Sci/Spec Premier wide mouth HDPE bottles. These bottles have been routinely used for fluorochemical sample Page 9 o f 65 E05-0210 Interim Report#13 Tennessee River Page 100 of 160 3M Environm ental Laboratory E05-0210 Interim Report#13 Exygen Protocol Number: P0001 131 collection at the testing facility and have been shown to be free of PFBS, PFHS and PFOS. Samples will be added to each container to a volumetric fill line at 200 mL. A field duplicate, a low field spike and a high field spike of each sample will be collected. The low and high field spike bottles will contain PFBS, PFHS and PFOS as well as perfluorooctanoic acid (PFOA) and 1.2-13C perfluorooctanoic acid (13C PFOA). PFOA and 13C PFOA are included in the solutions used to spike the samples. The results for PFOA and 13C PFOA will not be reported in this study. Exygen will supply one field blank (control water) and two field blank spikes (control water fortified with PFBS, PFHS and PFOS at a low and high level) for every twenty samples collected. At the testing facility, each water sample (excluding field duplicates and field spikes) will be extracted in duplicate and will also be fortified at a low and high concentration with PFBS, PFHS and PFOS and processed through the described procedure to determine method accuracy and to check for bias. For soil, sediment, clams, and vegetation, Exygen will supply one 500 mL precleaned Sci/Spec Premier wide mouth HDPE bottle per sample collected or a zip-seal bag. All containers/bags used for sample collection will be shipped to the sample location. Samples will be added to each container or bag in the field. At the testing facility, each sample will be extracted in duplicate and will also be fortified at a known concentration with PFBS, PFHS and PFOS at both a low and high level and processed through the described procedure to determine method accuracy and to check for bias. For small mammal liver, Exygen will supply a 50 mL polypropylene centrifuge tube. For small mammal serum, Exygen will supply a collection kit for each sample containing serum separator tubes (red top), vacutainers, needle holders and needles, transfer pipettes, and polypropylene tubes. At the testing facility, each liver and serum sample will be extracted in duplicate and will also be fortified at a known concentration with PFBS, PFHS and PFOS at both a low and high level and processed through the described procedure to determine method accuracy and to check for bias. Low and high spiking levels for each matrix are defined below: Matrix Low Spiking Level High Spiking Level Water 500 ng/L 5000 ng/L Soil 4 ng/g 40 ng/g Sediment 4 ng/g 40 ng/g Fish 10 ng/g 100 ng/g Clams 10 ng/g 100 ng/g Vegetation 10 ng/g 100 ng/g Small Mammal Liver 10 ng/g 100 ng/g Small Mammal Serum 10 ng/mL 100 ng/mL E05-0210 Interim Report#13 Tennessee River Page 10 o f 65 Page 101 of 160 3M Environm ental Laboratory E05-0210 Interim Report#13 Exygen Protocol Number: P 0001131 Recoveries are anticipated to be between 70% and 130% of the fortified levels; however, the exact precision and accuracy will be determined by the analysis of the quality control samples described above. A statement of accuracy will be included in the final report. M E T H O D FO R C O N TR O L OF BIAS Control of bias will be addressed by taking representative sub-samples from a homogeneous mixture of each matrix from untreated control samples, and by analyzing at least two levels of fortifications. S TA TIS TIC A L M ETH O D S Statistics will be limited to those specified in the subject methods and to the calculation of average recoveries, as applicable. GLP STATEMENT All aspects of this study shall be performed and reported in compliance with EPA TSCA Good Laboratory Practice Standards 40 CFR 792. The final report or data package (supplied to the Sponsor) shall contain a statement that the study was conducted in compliance with current and applicable GLP standards and will outline any deviations in the study from those standards. This statement will be signed by the Study Director and Sponsor Representative. REPORT A final report will be prepared by the principal investigator or their designee at the conclusion of the study. The report will include, but will not be limited to, the following: The name and address of the Study Director, Sponsor Representative, and of the testing facility. A statement of GLP compliance (any related documentation, such as chain-of-custody records, must be in the study records). E05-0210 Interim Report#13 Tennessee River Page / / o f 65 Page 102 of 160 3M Environm ental Laboratory E05-0210 Interim Report#13 Exygen Protocol Number: P 0001131 The signed and dated statement by the Exygen Research Quality Assurance Unit regarding dates of study inspections and dates findings were reported to the Study Director and Management. A description of the exact analytical conditions employed in the study. If the subject method was followed exactly, it is necessary to include only a copy of the analytical method. Any modifications to this method will be incorporated into the report. If the method is photo-reduced, the project number and page number must be included on each page. Description of the instrumentation used and operating conditions. All results from all sets analyzed. Control and fortified samples will be identified and the data table will include sample number and fortification level. Representative chromatograms for each analyte in each matrix, including chromatograms of a standard and a control sample, and a chromatogram at a fortification level. The location of the analyte peaks will be clearly identified in all chromatograms. All circumstances that may have affected the quality or integrity of the data will be documented in the report. Locations where raw data and the final report are to be archived. Additions or corrections to the final report shall be in the form of an amendment signed by the Study Director. The amendment shall clearly identify that part of the report that is being altered and the reasons for the alterations. The amendment will be signed and dated by the Study Director and the Sponsor Representative. All applicable requirements for reporting of study results as per 40 CFR 792.185. SAFETY AND HEALTH Laboratory personnel will practice good sanitation and health habits. Every reasonable precaution shall be taken to prevent inadvertent exposure of personnel and the environment to the test or reference substance(s). E05-0210 Interim Report#13 Tennessee River Page 12 o f 65 Page 103 of 160 3M Environm ental Laboratory E05-0210 Interim Report#13 Exygen Protocol Number: P 0001131 AMENDMENTS TO PROTOCOL All significant changes to the analytical protocol outlined here will be expressed in writing, signed and dated by the Study Director and Sponsor Representative. Amendments usually will be issued prior to initiation of study plan change. However, when a change is required without sufficient time for the issue of a written amendment, that change may be effected verbally with supporting documentation signed and dated by the Study Director and followed with a written amendment as soon as possible. In this case, the effective date of the written amendment will be the date of the documented change. Copies of the signed amendments will be appended to all distributed study plan copies. The original amendment will be maintained with the original study plan. Any deviations from the study plan or from the analytical method as provided will be documented and reported promptly to the Sponsor Representative. D A TA RECORD K E EP IN G Records to be maintained include the following (as appropriate): Sample tracking sheet(s) Sample receipt records, storage history, and chains of custody History and preparation of standards (stock, fortification, calibration) Description of any modifications to the method Instrument run sheets, bench-sheets or logs Analytical data tables All chromatographic and instrumental conditions Sample extraction and analysis dates A complete listing of study personnel, signatures and initials Chronological presentation of all study correspondence Any other documentation necessary for the reconstruction of the study Chromatograms- All chromatograms will contain the following: Sample identification, injection date, arrow or other indication of the area of interest, and injection number corresponding to the run. Additionally, fortifications will include the amount of analyte added and the sample number of the sample that was fortified. Analytical standard chromatograms will additionally include the concentration (e.g., pg/mL). Page 13 o f 65 E05-0210 Interim Report#13 Tennessee River Page 104 of 160 3M Environm ental Laboratory E05-0210 Interim Report#13 Exygen Protocol Number: P 0001131 As part of the documentation the following sheets will be included in each analytical set: a run sheet listing the samples to be run in the set, and an instrument conditions sheet describing the instrument type and operating conditions. Q U A L IT Y ASSURANCE The QA Unit of Exygen Research will inspect the study at intervals adequate to assure compliance with GLP's, and will report the findings of audits to the Study Director, Exygen Management, and the Sponsor Representative. R E TE N TIO N OF D A TA AND A R C H IV IN G All hard copy raw data, including, but not limited to, the original chromatograms, worksheets, correspondence, and results shall be included with the data package submitted to the Study Director. These will be archived with the original study plan, amendments, final report, and all pertinent information from the Sponsor. The testing facility shall keep all electronic raw data and any instrument, equipment, and storage logs for the period of time specified in 40 CFR 792.195. An exact copy of the materials submitted to the study director will also be kept at Exygen Research. Exygen will obtain permission from the study director before discarding or returning samples. E05-0210 Interim Report#13 Tennessee River Page 14 o f 65 Page 105 of 160 3M Environm ental Laboratory E05-0210 Interim Report#13 Exygen Protocol Number: P 0001131 APPENDIX I ANALYTICAL METHODS V0001780: "Method of Analysis for the Determination of Perfluorooctanoic Acid (PFOA) in Water by LC/MS/MS" V0001781: "Method of Analysis for the Determination of Perfluorooctanoic Acid (PFOA) in Soil by LC/MS/MS" V0001782: "Method of Analysis for the Determination of Perfluorooctanoic Acid (PFOA) in Sediment by LC/MS/MS" V0001783: "Method of Analysis for the Determination of Perfluorooctanoic Acid (PFOA) in Fish and Clams by LC/MS/MS" V0001784: "Method of Analysis for the Determination of Perfluorooctanoic Acid (PFOA) in Vegetation by LC/MS/MS" V0001785: "Method of Analysis for the Determination of Perfluorooctanoic Acid (PFOA) in Small Mammal Liver by LC/MS/MS" V0001786: "Method of Analysis for the Determination of Perfluorooctanoic Acid (PFOA) in Small Mammal Serum by LC/MS/MS" E05-0210 Interim Report#13 Tennessee River Page 15 o f 65 Page 106 of 160 3M Environm ental Laboratory E05-0210 Interim Report#13 Exygen Protocol Number: P0001131 ANALYTICAL METHOD Method Number: V0001780 Method of Analysis for the Determination of Perfluorooctanoic Acid (PFOA) in Water by LC/MS/MS Analytical Testing Facility: Exygen Research 3058 Research Drive State College, PA 16801 Approved By: c Paul Connolly Technical Leader, LC-MS, Exygen Research 7,1fl?fI*X / lohn Flaherty / ' VV iicroe PDrr<easoiidfleinntt, OHnpperriations, Exygen Research <0)zA/oii Date Date Total Pages: 7 E05-0210 Interim Report#13 Tennessee River Page 16 o f 65 Page 107 of 160 3M Environm ental Laboratory E05-0210 Interim Report#13 Exygen Protocol Number: P 0001131 Exygen Research Method Number V0001780 | ANALYTICAL METHOD Method o f Analysis for the Determination o f Perfluorooctanoic Acid (PFOA) in Water by LC/MS/MS 1.0 Scope This method is to be employed for the isolation and quantitation of perfluorooctanoic acid by High Performance Liquid Chromatography coupled to a tandem Mass Spectrometric Detector (LC/MS/MS) in water. 2.0 Safety 2.1 Always observe safe laboratory practices. 2.2 Consult the appropriate M SDS before handling any chemical for proper safety precautions. 3.0 Sample Requirement 3.1 At least 40 mL o f test sample for extraction. 3.2 No sample processing is needed for water samples. 3.3 Samples stored refrigerated should be allowed to equilibrate to room temperature. 3.4 All samples must be thoroughly mixed before being sampled for extraction. 3.5 Any samples containing particles should be centrifuged at -3000 rpm for -5 minutes and the supernatant used for the extraction. 3.6 Sample collection procedures will be specified in the sampling plan for this project. 4.0 Reagents and Standards 4.1 Water - HPLC grade 4.2 Methanol - HPLC grade 4.3 Ammonium Acetate - A.C.S. Reagent Grade 4.4 Perfluorooctanoic Acid - Sigma-Aldrich 5.0 Instrument and Equipment 5.1 A high performance liquid chromatograph capable o f pumping up to 2 solvents equipped with a variable volume injector capable of injecting 5-200 pL connected to a tandem Mass Spectrometer (LC/MS/M S). 5.2 A device to collect raw data for peak integration and quantitation. 5.3 Analytical balance capable of reading to 0.00001 g. 5.4 50 mL disposable polypropylene centrifuge tubes. 5.5 15 mL disposable polypropylene centrifuge tubes. 5.6 Disposable micropipets (50-1 OOuL, 100-200uL). 5.7 125-mLLDPE narrow-mouth bottles. 5.8 2 mL clear HPLC vial kit. 5.9 Disposable pipettes. 5.10 Autopipettes (100-1000 pL and 10-100 pL), with disposable tips. 5.11 Waters Sep Pak Vac 6 cc (lg ) tC18 SPE cartridges. Page 2 o f 7 E05-0210 Interim Report#13 Tennessee River Page 17 o f 65 Page 108 of 160 3M Environm ental Laboratory E05-0210 Interim Report#13 Exygen Protocol Number: P 0001131 Exygen Research Method Number V0001780 ANALYTICAL METHOD Method o f Analysis for the Determination o f Perfluorooctanoic Acid (PFOA) in Water bv LC /M S/M S ' 5.12 SPE vacuum manifold. 5.13 Centrifuge capable o f spinning 50 mL polypropylene tubes at 3000 rpm. 6.0 Chromatographic System 6.1 Analytical Column: Fluophase RP (Keystone Scientific), 2.1 mm x 50 mm, 5p (P/N: 82505-052130) 6.2 Temperature: 30C 6.3 Mobile Phase (A) : 2 mM Ammonium Acetate in Water 6.4 Mobile Phase (B) : Methanol 6.5 Gradient Program: Time (min) 0.0 1.0 8.0 20.0 22.5 %A 65 65 25 25 65 Flow Rate % B (mL/min) 35 0.3 35 0.3 75 0.3 75 0.3 35 0.3 6.6 Injection Volume: 15 pL (can be increased to as much as 50 pL). 6.7 Quantitation: Peak Area - external standard calibration curve. 6.8 Run Time: ~ 23 minutes. The above conditions are intended as a guide and may be changed in order to optimize the HPLC system. 7.0 MS/MS System 7.1 Mode: Electrospray Negative MRM mode, monitoring 4 1 3 - 369 m/z. The above conditions are intended as a guide and may be changed in order to optimize the MSMS system. 8.0 Preparation o f Solutions 8.1 Mobile Phase 8.1.1 2 mM ammonium acetate in water is prepared by adding 0.154 g of ammonium acetate to 1000 mL o f water. Alternate volumes may be prepared. Page 3 of ' E05-0210 Interim Report#13 Tennessee River Page 18 o f 65 Page 109 of 160 3M Environm ental Laboratory E05-0210 Interim Report#13 Exygen Protocol Number: P0001131 Exygen Research Method Number V0001780 | ANALYTICAL METHOD Method o f Analysis for the Determination o f Perfluorooctanoic Acid (PFOA) in Water by LC /M S/M S 9.0 Standard Preparation 9.1 Standard Stock/Fortification Solution 9.1.1 Prepare a stock solution o f ~100 pg/mL o f PFOA by weighing 10 mg of analytical standard (corrected for purity) and dilute to 100 mL with methanol in a 125-mL LDPE bottle. 9.1.2 A 10 pg/mL fortification solution o f PFOA is prepared by bringing 10 mL o f the 100 pg/mL solution to a final volume o f 100 with methanol in a 125 mL LDPE bottle. 9.1.3 A 1.0 pg/mL fortification solution o f PFOA is prepared by bringing 10 mL o f the 10 pg/mL solution to a final volume of 100 with methanol m a 125 mL LDPE bottle. 9.1.4 A 0.1 pg/mL fortification solution o f PFOA is prepared by bringing 10 mL of the 1.0 pg/mL solution to a final volume o f 100 with methanol in a 125 mL LDPE bottle. 9.1.5 A 0.01 pg/mL fortification solution o f PFOA is prepared by bringing 10 mL o f the 0.1 pg/mL solution to a final volume of 100 with methanol in a 125 mL LDPE bottle. 9.1.6 The stock and fortification solutions are to be stored in a refrigerator at approximately 4C and are stable for a maximum period of 6 months from the date o f preparation. 9.2 Standard Calibration Solutions 9.2.1 9.2.2 LC/MS/MS calibration standards are prepared in HPLC water. The calibration standards are processed through the extraction procedure, identical to samples. The following is a typical example: additional concentrations may be prepared as needed. Final Concentration Fortification Volume of Concentration of Calibration o f Fortification Volume Fortified Control Calibration Standard ID Solution (ppb) (PL) Sample (mL) Standard (ppt)* (example) 0 0 40 0 XCmmddyy-0 10 100 40 25 XCmmddyy-1 10 200 40 50 XCmmddyy-2 10 400 40 100 XCmmddyy-3 100 100 40 250 XCmmddyy-4 100 200 40 500 XCmmddyy-5 100 400 40 1000 XCmmddyy-6 * The extracted concentration o f the calibration standard is equal to 8x its initial concentration, due to the concentration o f the standard during the extraction (SPE). XC = extracted calibration standard. Page 4 of ^ E05-0210 Interim Report#13 Tennessee River Page 19 o f 65 Page 110 of 160 3M Environm ental Laboratory E05-0210 Interim Report#13 Exygen Protocol Number: P0001131 Exygen Research Method Number V0001780 ANALYTICAL METHOD Method o f Analysis for the Determination o f Perfluorooctanoic Acid (PFOA) in Water by LC/MS/MS 9.2.3 9.2.4 9.2.5 A zero standard solution (reagent blank) must be prepared with each set o f standards extracted. Store all extracted calibration standards in 15-mL polypropylene tubes at 2C to 6C, up to two weeks. Alternate volumes and concentrations o f standards may be prepared as needed. 10.0 Batch Set Up 10.1 Each batch o f samples extracted (typically 20 or less) must include at least one reagent control (method blank using HPLC water) and two reagent controls fortified at known concentrations (lab control spike) to verify procedural recovery for the batch. 10.2 Requirements for field and laboratory duplicates and spikes will be specified in the quality assurance plan for this project. 11.0 Sample Extraction 11.1 Measure 40 mL o f sample or a portion o f sample diluted to 40 mL with water into 50 mL polypropylene centrifuge tubes (fortify as needed, replace lid and mix well). 11.2 Condition the Cig SPE cartridges (1 g, 6 mL) by passing 10 mL methanol followed by 5 mL o f HPLC water (~ 2 drop/sec). Do not let column run dry 11.3 Load sample on conditioned Cig SPE cartridge. Discard eluate. 11.4 Elute with ~5 mL 100% methanol. Collect 5 mL o f eluate into graduated 15 mL polypropylene centrifuge tubes (final volume = 5 mL). 11.5 Analyze samples using electrospray LC/MS/MS. 12.0 Chromatography 12.1 Inject the same amount o f each standard, sample and fortified sample into the LC/MS/MS system. A calibration standard must precede and follow all analyzed samples. 12.2 Standards o f PFOA corresponding to at least five or more concentration levels must be included in an analytical set. 12.3 An entire set o f extracted calibration standards must be included at the beginning and at the end o f a sample set. Extracted standards must be interspersed between every 5-10 samples. As an alternative, an entire set of extracted calibration standards may be injected at the beginning of a set followed by extracted calibration standards interspersed every 5-10 samples (to account for a second set o f extracted standards). In either case, extracted calibration standards must be the first and last injection in a sample set 12.4 Use linear standard curves for quantitation. Linear standard curves are generated for the analyte by linear regression using 1/x weighting of peak area Page 5 of E05-0210 Interim Report#13 Tennessee River Page 20 o f 65 Page 111 of 160 3M Environm ental Laboratory E05-0210 Interim Report#13 Exygen Protocol Number: P0001131 Exygen Research Method Number V0001780 | ANALYTICAL METHOD Method o f Analysis for the Determination o f Perfluorooctanoic Acid (PFOA) in Water by LC/MS/MS versus calibration standard concentration using MassLynx 3.3 (or equivalent) software system. 12.5 Sample response should not exceed standard responses. Any samples that exceed standard responses should be further diluted and reanalyzed. 13.0 Acceptance Criteria 13.1 Chromatogram must show a peak of a daughter ion at 369 amu from a parent o f 413 amu. The 413 amu parent corresponds to the PFOA anion, while the daughter ion (369 amu) represents the loss of carbon dioxide 13.2 Method blanks must not contain PFOA at levels greater than the LOQ. If a blank contains PFOA at levels greater than 50 ng/L, then a new blank sample must be obtained and the entire set must be re-extracted. 13.3 Recoveries o f control spikes and matrix spikes must be between 70-130% of their known values. If a control spike falls outside the acceptable limits, the entire set o f samples should be re-extracted. Any matrix spike outside 70 130% should be evaluated by the analyst to determine if re-extraction is warranted. 13.4 Any calibration standard found to be a statistical outlier by using the Huge Error Test, may be excluded from the calculation o f the calibration curve. However, the total number o f extracted calibration standards that could be excluded must not exceed 20% o f the total number o f extracted standards injected. 13.5 The correlation coefficient (R) for calibration curves generated must be >0.992 (R2 >0.985). If calibration results fall outside these limits, then appropriate steps must be taken to adjust instrument operation, and the standards or the relevant set o f samples should be reanalyzed. 13.6 Retention times between standards and samples must not drift more than 4 % within an analytical run. If retention time drift exceeds this limit within an analytical run then the set must be reanalyzed. 14.0 Calculations 14.1 Use the following equation to calculate the amount o f PFOA found (in ng/L, based on peak area) using the standard curve (linear regression parameters) generated by the Mass Lynx software program: PFOA found (ng/L) = (Peak area - intercept) x DF slope DF = factor by which the final volume was diluted, if necessary. Page 6 of 7 E05-0210 Interim Report#13 Tennessee River Page 21 o f 65 Page 112 of 160 3M Environm ental Laboratory E05-0210 Interim Report#13 Exygen Protocol Number: P0001131 Exygen Research Method Number V0001780 | ANALYTICAL METHOD Method o f Analysis for the Determination o f Perfluorooctanoic Acid (PFOA) in Water by LC/MS/MS 14.2 For samples fortified with known amounts o f PFOA prior to extraction, use the following equation to calculate the percent recovery. Recovery (%) = [ total analyte found (ng/L) - analyte found in control (ng/L)] ;1QQ analyte added (ng/L) Page 7 o f ' E05-0210 Interim Report#13 Tennessee River Page 22 o f 65 Page 113 of 160 3M Environm ental Laboratory E05-0210 Interim Report#13 Exygen Protocol Number: P 0001131 ANALYTICAL METHOD Method Number: V0001781 Method of Analysis for the Determination of Perfluorooctanoic Acid (PFOA) in Soil by LC/MS/MS Analytical Testing Facility: Exygen Research 3058 Research Drive State College, PA 16801 Approved By: Paul Connolly Technical Leader, LC-MS, Exygen Research I Date Total Pages: 7 E05-0210 Interim Report#13 Tennessee River Page 23 o f 65 Page 114 of 160 3M Environm ental Laboratory E05-0210 Interim Report#13 Exygen Protocol Number: P 0001131 Exygen Research Method Number V0001781 1 ANALYTICAL METHOD Method o f Analysis for the Determination o f Perfluorooctanoic Acid (PFOA) in Soil by LC/MS/MS 1.0 Scope This method is to be employed for the isolation and quantitation o f perfluorooctanoic acid by High Performance Liquid Chromatography coupled to a tandem Mass Spectrometric Detector (LC/MS/MS) in soil. 2.0 Safety 2.1 Always observe safe laboratory practices. 2.2 Consult the appropriate M SDS before handling any chemical for proper safety precautions. 3.0 Sample Requirement 3.1 At least 15 g o f test sample for extraction. 3.2 No sample processing is needed for soil samples. 3.3 Samples stored refrigerated should be allowed to equilibrate to room temperature. 3.4 All samples must be thoroughly mixed before being sampled for extraction. 3.5 Sample collection procedures will be specified in the sampling plan for this project. 4.0 Reagents and Standards 4.1 Water - HPLC grade 4.2 Methanol - HPLC grade 4.3 Ammonium Acetate - A.C.S. Reagent Grade 4.4 Perfluorooctanoic Acid - Sigma-Aldrich 5.0 Instrument and Equipment 5.1 A high performance liquid chromatograph capable o f pumping up to 2 solvents equipped with a variable volume injector capable of injecting 5-200 pL connected to a tandem Mass Spectrometer (LC/MS/M S). 5.2 A device to collect raw data for peak integration and quantitation. 5.3 Analytical balance capable o f reading to 0.00001 g. 5.4 50 mL disposable polypropylene centrifuge tubes. 5.5 15 mL disposable polypropylene centrifuge tubes. 5.6 Disposable micropipets (50-1 OOuL, 100-200uL). 5.7 125-mL LDPE narrow-mouth bottles. 5.8 2 mL clear HPLC vial kit. 5.9 Disposable pipettes. 5.10 Autopipettes (100-1000 pL and 10-100 pL), with disposable tips. 5.11 Waters Sep Pak Vac 6 cc (lg ) tC18 SPE cartridges. 5.12 SPE vacuum manifold. 5.13 Ultrasonic bath. Page 2 o f 7 E05-0210 Interim Report#13 Tennessee River Page 24 o f 65 Page 115 of 160 3M Environm ental Laboratory E05-0210 Interim Report#13 Exygen Protocol Number: P 0001131 Exygen Research Method Number V0001781 | ANALYTICAL METHOD Method o f Analysis for the Determination o f Perfluorooctanoic Acid (PFOA) in Soil by LC /M S/M S 5.14 Wrist-action shaker. 5.15 Centrifuge capable of spinning 50 mL polypropylene tubes at 5000 rpm. 6.0 Chromatographic System 6.1 Analytical Column: Fluophase RP (Keystone Scientific), 2.1 mm x 50 mm, 5p (P/N: 82505-052130) 6.2 Temperature: 30C 6.3 Mobile Phase (A) : 2 mM Ammonium Acetate in Water 6.4 Mobile Phase (B) : Methanol 6.5 Gradient Program: Time (min) 0.0 1.0 8.0 20.0 22.5 %A 65 65 25 25 65 Flow Rate % B (mL/min) 35 0.3 35 0.3 75 0.3 75 0.3 35 0.3 6.6 Injection Volume: 15 pL (can be increased to as much as 50 pL). 6.7 Quantitation: Peak Area - external standard calibration curve. 6.8 Run Time: ~ 23 minutes. The above conditions are intended as a guide and may be changed in order to optimize the HPLC system. 7.0 MS/MS System 7.1 Mode: Electrospray Negative MRM mode, monitoring 413 369 m/'z for PFOA. The above conditions are intended as a guide and may be changed in order to optimize the MSMS system. 8.0 Preparation of Solutions 8.1 Mobile Phase 8.1.1 2 mM ammonium acetate in water is prepared by adding 0.154 g of ammonium acetate to 1000 mL of water. Alternate volumes may be prepared. Page 3 of 7 E05-0210 Interim Report#13 Tennessee River Page 25 o f 65 Page 116 of 160 3M Environm ental Laboratory E05-0210 Interim Report#13 Exygen Protocol Number: P 0001131 Exygen Research M eth od N u m ber V 0 0 0 1781 ANALYTICAL METHOD Method o f Analysis for the Determination o f Perfluorooctanoic Acid (PFOA) in Soil by LC/MS/MS 9.0 Standard Preparation 9.1 Standard Stock/Fortification Solution 9.1.1 Prepare a stock solution o f -100 pg/mL o f PFOA by weighing 10 mg o f analytical standard (corrected for purity) and dilute to 100 mL with methanol in a 125-mL LDPE bottle. 9.1.2 A 10 pg/mL fortification solution o f PFOA is prepared by bringing 10 mL o f the 100 pg/mL solution to a final volume o f 100 with methanol in a 125 mL LDPE bottle. 9.1.3 A 1.0 pg/mL fortification solution o f PFOA is prepared by bringing 10 mL o f the 10 pg/mL solution to a final volume o f 1 0 0 w it h m e th a n o l m a 125 mL LDPE bottle. 9.1.4 A 0.1 pg/mL fortification solution o f PFOA is prepared by bringing 10 mL o f Jhe 1.0 pg/mL solution to a final volume o f 100 with methanol in a 125 mL LDPE bottle. 9.1.5 A 0.01 pg/mL fortification solution o f PFOA is prepared by bringing 10 mL o f the 0.1 pg/mL solution to a final volume of 100 with methanol in a 125 mL LDPE bottle. 9.1.6 The stock and fortification solutions are to be stored in a refrigerator at approximately 4C and are stable for a maximum period of 6 months from the date o f preparation. 9.2 Standard Calibration Solutions 9.2.1 9.2.2 LC/MS/MS calibration standards are prepared in HPLC water. The calibration standards are processed through the extraction procedure, identical to samples. The following is a typical example: additional concentrations may be prepared as needed. Final Concentration Fortification Volume of Concentration of Calibration o f Fortification Volume Fortified Control Calibration Standard ID Solution (ppb) (PL) Sample (mL) Standard (ppt)* (example) 0 0 40 0 XCmmddyy-0 10 100 40 25 XCmmddyy-1 10 200 40 50 XCmmddyy-2 10 400 40 100 XCmmddyy-3 100 100 40 250 XCmmddyy-4 100 200 40 500 XCmmddyy-5 100 400 40 1000 XCmmddyy-6 * The extracted concentration o f the calibration standard is equal to 8x its initial concentration, due to the concentration o f the standard during the extraction (SPE). XC = extracted calibration standard. P age 4 of 7 E05-0210 Interim Report#13 Tennessee River Page 26 o f 65 Page 117 of 160 3M Environm ental Laboratory E05-0210 Interim Report#13 Exygen Protocol Number: P 0001131 Exygen Research Method Number V0001781 | ANALYTICAL METHOD Method o f Analysis for the Determination o f Perfluorooctanoic Acid (PFOA) in Soil by LC/MS/MS 9.2.3 9.2.4 9.2.5 A zero standard solution (reagent blank) must be prepared with each set o f standards extracted. Store all extracted calibration standards in 15-mL polypropylene tubes at 2C to 6C, up to two weeks. Alternate volumes and concentrations o f standards may be prepared as needed. 10.0 Batch Set Up 10.1 Each batch o f samples extracted (typically 20 or less) must include at least one reagent control (method blank using 5 mL o f methanol) and two reagent controls fortified at known concentrations (lab control spike) to verify procedural recovery for the batch. 10.2 Requirements for field and laboratory duplicates and spikes will be specified in the quality assurance plan for this project. 11.0 Sample Extraction 11.1 Weigh 5 g o f sample into 50 mL polypropylene centrifuge tubes (fortify as needed, replace lid and mix well). 11.2 Add 5 mL o f methanol and shake on a wrist action shaker for ~15 minutes. 11.3 Transfer the tubes to an ultrasonic bath and sonicate for -15 minutes. 11.4 Bring the volume up to 40 mL with water in the 50 mL polypropylene centrifuge tube. 11.5 Centrifuge for -1 0 minutes at -3000 rpm. 11.6 Condition the Cig SPE cartridges (1 g, 6 mL) by passing 10 mL methanol followed by 5 mL o f HPLC water (~ 2 drop/sec). Do not let column run dry 11.7 Load (decant) the sample on the conditioned C )8 SPE cartridge. Discard eluate. 11.8 Elute with -5 mL 100% methanol. Collect 5 mL o f eluate into graduated 15 mL polypropylene centrifuge tubes (final volume = 5 mL). 11.9 Analyze samples using electrospray LC/MS/MS. 12.0 Chromatography 12.1 Inject the same amount o f each standard, sample and fortified sample into the LC/MS/MS system. A calibration standard must precede and follow all analyzed samples. 12.2 Standards o f PFOA corresponding to at least five or more concentration levels must be included in an analytical set. 12.3 An entire set o f extracted calibration standards must be included at the beginning and at the end o f a sample set. Extracted standards must be interspersed between every 5-10 samples. As an alternative, an entire set of Page 5 of 7 E05-0210 Interim Report#13 Tennessee River Page 27 o f 65 Page 118 of 160 3M Environm ental Laboratory E05-0210 Interim Report#13 Exygen Protocol Number: P 0001131 Exygen Research Method Number V0001781 P ANALYTICAL METHOD Method o f Analysis for the Determination o f Perfluorooctanoic Acid (PFOA) in Soil by LC /M S/M S ' extracted calibration standards may be injected at the beginning of a set followed by extracted calibration standards interspersed every 5-10 samples (to account for a second set o f extracted standards). In either case, extracted calibration standards must be the first and last injection in a sample set. 12.4 Use linear standard curves for quantitation. Linear standard curves arc generated for the analyte by linear regression using 1/x weighting of peak area versus calibration standard concentration using MassLynx 3.3 (or equivalent) software system. 12.5 Sample response should not exceed standard responses. Any samples that exceed standard responses should be further diluted and reanalyzed. 13.0 Acceptance Criteria 13.1 Chromatogram must show a peak o f a daughter ion at 369 amu from a parent of 413 amu. The 413 amu parent corresponds to the PFOA anion, while the daughter ion (369 amu) represents the loss o f carbon dioxide. 13.2 Method blanks must not contain PFOA at levels greater than the LOQ. If a blank contains PFOA at levels greater than 50 ng/L, then a new blank sample must be obtained and the entire set must be re-extracted. 13.3 Recoveries o f control spikes and matrix spikes must be between 70-130% of their known values. If a control spike falls outside the acceptable limits, the entire set o f samples should be re-extracted. Any matrix spike outside 70 130% should be evaluated by the analyst to determine if re-extraction is warranted. 13.4 Any calibration standard found to be a statistical outlier by using the Huge Error Test, may be excluded from the calculation o f the calibration curve. However, the total number o f extracted calibration standards that could be excluded must not exceed 20% o f the total number o f extracted standards injected. 13.5 The correlation coefficient (R) for calibration curves generated must be >0.992 (R2 0.985). If calibration results fall outside these limits, then appropriate steps must be taken to adjust instrument operation, and the standards or the relevant set o f samples should be reanalyzed. 13.6 Retention times between standards and samples must not drift more than 4 % within an analytical run. If retention time drift exceeds this limit within an analytical run then the set must be reanalyzed. Page 6 of 7 E05-0210 Interim Report#13 Tennessee River Page 28 o f 65 Page 119 of 160 3M Environm ental Laboratory E05-0210 Interim Report#13 Exygen Protocol Number: P 0001131 Exygen Research Method NumberV0001781 ANALYTICAL METHOD Method o f Analysis for the Determination o f Perfluorooctanoic Acid (PFOA) in Soil bv LC/MS/MS ' \ 14.0 Calculations 14.1 Use the following equation to calculate the amount o f PFOA found (in ng/L, based on peak area) using the standard curve (linear regression parameters) generated by the Mass Lynx software program: PFOA found (ng/L) = (Peak area - intercept) x DF slope DF = factor by which the final volume was diluted, if necessary. 14.2 For samples fortified with known amounts o f PFOA prior to extraction, use the following equation to calculate the percent recovery. Recovery (%) = [ total analyte found (ng/L) - analyte found in control (ng/L)] ^ analyte added (ng/L) 14.3 Use the following equation to convert the amount o f PFOA found in ng/L to ng/g (ppb). PFOA found (ppb) = fPFOA found (ng/L) x volume extracted t'0.04L)l sample weight (5 g) 14.4 Use the following equation to calculate the amount o f PFOA found in ppb based on dry weight. PFOA found (ppb) dry weight = PFOA found (ppb) x [100% / total solids(%)] Page 7 of 7 E05-0210 Interim Report#13 Tennessee River Page 29 o f 65 Page 120 of 160 3M Environm ental Laboratory E05-0210 Interim Report#13 Exygen Protocol Number: P 0001131 ANALYTICAL METHOD Method Number: V0001782 Method of Analysis for the Determination of Perfluorooctanoic Acid (PFOA) in Sediment by LC/MS/MS Analytical Testing Facility: Exygen Research 3058 Research Drive State College, PA 16801 Approved By: C -jL ____ Paul Connolly I Technical Leader, LC-MS, Exygen Research j/n f b l/ ohn Flaherty Vice President, Operations, Exygen Research ___ lo lW o '/ Date Date Total Pages: 7 E05-0210 Interim Report#13 Tennessee River Page 30 o f 65 Page 121 of 160 3M Environm ental Laboratory E05-0210 Interim Report#13 Exygen Protocol Number: P 0001131 Exygen Research Method Number V0001782 | ANALYTICAL METHOD j Method of Analysis for the Determination o f Perfluorooctanoic Acid (PFOA) in Sediment bv LC/MS/MS ' 1.0 Scope This method is to be employed for the isolation and quantitation of perfluorooctanoic acid by High Performance Liquid Chromatography coupled to a tandem Mass Spectrometric Detector (LC/MS/MS) in sediment. 2.0 Safety 2.1 Always observe safe laboratory practices. 2.2 Consult the appropriate M SDS before handling any chemical for proper safety precautions. 3.0 Sample Requirement 3.1 At least 30 g o f test sample for extraction. 3.2 No sample processing is needed for sediment samples. 3.3 Samples stored refrigerated should be allowed to equilibrate to room temperature. 3.4 All samples must be thoroughly mixed before being sampled for extraction. 3.5 Sample collection procedures will be specified in the sampling plan for this project. 4.0 Reagents and Standards 4.1 Water - HPLC grade 4.2 Methanol - HPLC grade 4.3 Acetic Acid - Reagent grade 4.4 Ammonium Acetate - A.C.S. Reagent Grade 4.5 Perfluorooctanoic Acid - Sigma-Aldrich 5.0 Instrument and Equipment 5.1 A high performance liquid chromatograph capable o f pumping up to 2 solvents equipped with a variable volume injector capable o f injecting 5-200 pL connected to a tandem Mass Spectrometer (LC/MS/M S). 5.2 A device to collect raw data for peak integration and quantitation. 5.3 Analytical balance capable o f reading to 0.00001 g. 5.4 50 mL disposable polypropylene centrifuge tubes. 5.5 15 mL disposable polypropylene centrifuge tubes. 5.6 Disposable micropipets (50-1 OOuL, 100-200uL). 5.7 125-mL LDPE narrow-mouth bottles. 5.8 2 mL clear HPLC vial kit. 5.9 Disposable pipettes. 5.10 Autopipettes (100-1000 pL and 10-100 pL), with disposable tips. 5.11 Waters Sep Pak Vac 6 cc (lg ) tC18 SPE cartridges. 5.12 SPE vacuum manifold. Page 2 of 7 E05-0210 Interim Report#13 Tennessee River Page 31 o f 65 Page 122 of 160 3M Environm ental Laboratory E05-0210 Interim Report#13 Exygen Protocol Number: P 0001131 Exygen Research Method Number V0001782 ANALYTICAL METHOD \ Method of Analysis for the Determination of Perfluorooctanoic Acid (PFOA) in Sediment b\ LC/MS/MS ' 5.13 Vortexer. 5.14 Wrist-action shaker. 5.15 Centrifuge capable o f spinning 50 mL polypropylene tubes at 3000 rpm. 6.0 Chromatographic System 6.1 Analytical Column: Fluophase RP (Keystone Scientific), 2.1 mm x 50 mm, 5p (P/N: 82505-052130) 6.2 Temperature: 30C 6.3 Mobile Phase (A) : 2 mM Ammonium Acetate in Water 6.4 Mobile Phase (B) : Methanol 6.5 Gradient Program: Time (min) 0.0 1.0 8.0 20.0 22.5 %A 65 65 25 25 65 Flow Rate % B (mL/min) 35 0.3 35 0.3 75 0.3 75 0.3 35 0.3 6.6 Injection Volume: 15 pL (can be increased to as much as 50 pL). 6.7 Quantitation: Peak Area - external standard calibration curve. 6.8 Run Time: ~ 23 minutes. The above conditions are intended as a guide and may be changed in order to optimize the HPLC system. 7.0 MS/MS System 7.1 Mode: Electrospray Negative MRM mode, monitoring 413 - 369 m/z for PFOA. The above conditions are intended as a guide and may be changed in order to optimize the MSMS system. 8.0 Preparation o f Solutions 8.1 Mobile Phase 8.1.1 2 mM ammonium acetate in water is prepared by adding 0.154 g of ammonium acetate to 1000 mL o f water. Page 3 of 7 E05-0210 Interim Report#13 Tennessee River Page 32 o f 65 ...... P a g e 1 2 3 o f 1 6 0 3M Environm ental Laboratory E05-0210 Interim Report#13 Exygen Protocol Number: P0001 131 Exygen Research Method Number V0001782 ANALYTICAL METHOD Method o f Analysis for the Determination o f Perfluorooctanoic Acid (PFOA) in Sediment bv LC/MS/MS 8.2 Extraction Solutions 8.2.1 1% acetic acid in water is prepared by adding 10 mL o f acetic acid to 1000 mL o f water. Alternate volumes may be prepared. 9.0 Standard Preparation 9.1 Standard Stock/Fortification Solution 9.1.1 Prepare a stock solution o f -100 pg/mL o f PFOA by weighing 10 mg of analytical standard (corrected for purity) and dilute to 100 mL with methanol in a 125-mL LDPE bottle. 9.1.2 A 10 pg/mL fortification solution o f PFOA is prepared by bringing 10 mL o f the 100 pg/mL solution to a final volume o f 100 with methanol in a 125 mL LDPE bottle. 9.1.3 A 1.0 pg/mL fortification solution o f PFOA is prepared by bringing 10 mL o f the 10 pg/mL solution to a final volume o f 100 with methanol in a 125 mL LDPE bottle. 9.1.4 A 0.1 pg/mL fortification solution o f PFOA is prepared by bringing 10 mL o f the 1.0 pg/mL solution to a final volume o f 100 with methanol in a 125 mL LDPE bottle. 9.1.5 A 0.01 pg/mL fortification solution o f PFOA is prepared by bringing 10 mL o f the 0.1 pg/mL solution to a final volume o f 100 with methanol in a 125 mL LDPE bottle. 9.1.6 The stock and fortification solutions are to be stored in a refrigerator at approximately 4C and are stable for a maximum period of 6 months from the date o f preparation. 9.2 Standard Calibration Solutions 9.2.1 LC/MS/MS calibration standards are prepared in methanol via dilution of the 0.1 pg/mL fortification solution. 9.2.2 The following is a typical example: additional concentrations may be Concentration o f Fortification Solution (ng/mL) 100 100 100 10 5 2 Volume (mL) 10 5 2 10 10 10 Diluted to (mL) 100 100 100 100 100 100 Final Concentration (ng/mL) 10.0 5.0 2.0 1.0 0.5 0.2 Page 4 of : E05-0210 Interim Report#13 Tennessee River Page 33 o f 65 Page 124 of 160 3M Environm ental Laboratory E05-0210 Interim Report#13 Exygen Protocol Number: P0001131 Exygen Research Method Number V0001782 | ANALYTICAL METHOD 1 Method o f Analysis for the Determination o f Perfluorooctanoic Acid (PFOA) in Sediment by LC/MS/MS 9.2.3 9.2.4 Store all calibration standards in 125-mL LDPE narrow-mouth bottles at 2C to 6C, up to six months. Alternate volumes and concentrations o f standards may be prepared as needed. 10.0 Batch Set Up 10.1 Each batch o f samples extracted (typically 20 or less) must include at least one untreated control and two untreated controls fortified at known concentrations (lab control spike) to verify procedural recovery for the batch. 10.2 Requirements for field and laboratory duplicates and spikes will be specified in the quality assurance plan for this project. 11.0 Sample Extraction 11.1 Weigh 5 g o f sample into 50 mL polypropylene centrifuge tubes (fortify as needed, replace lid and mix well). 11.2 Add 35 mL o f 1% acetic acid, cap, vortex and shake on a wrist action shaker for ~60 minutes. 11.3 Centrifuge the tubes at -3000 rpm for -2 0 minutes. 11.4 Condition the Cig SPE cartridges (1 g, 6 mL) by passing 10 mL methanol followed by 20 mL o f HPLC water (~ 2 drop/sec). Do not let column run dry 11.5 Load (decant) the sample on the conditioned Cig SPE cartridge. Discard eluate. 11.6 Add 20 mL o f methanol to the sediment left in the bottom of the 50 mL centrifuge tube. Cap, vortex and shake on a wrist action shaker for -30 minutes. 11.7 Centrifuge the tubes at -3000 rpm for -2 0 minutes. 11.8 Decant the methanol onto the same SPE cartridge. Collect the eluate. 11.9 Wash the column with 4 mL o f methanol. Collect the eluate and add it to the eluate collected in step 11.8. 11.10 Condition a second Cig SPE cartridge (1 g, 6 mL) by passing 10 mL methanol followed by 20 mL o f HPLC water (~ 2 drop/sec). Do not let column run dry 11.11 Add the methanol to -200 mL o f water and load on the second conditioned SPE cartridge. 11.12 Elute with -5 mL 100% methanol. Collect 5 mL o f eluate into graduated 15 mL polypropylene centrifuge tubes (final volume = 5 mL). 11.13 Analyze samples using electrospray LC/M S/M S. Page 5 of 7 E05-0210 Interim Report#13 Tennessee River Page 34 o f 65 Page 125 of 160 3M Environm ental Laboratory E05-0210 Interim Report#13 Exygen Protocol Number: P0001131 Exygen Research Method Number V0001782 | ANALYTICAL METHOD [ Method o f Analysis for the Determination o f Perfluorooctanoic Acid (PFOA) in Sediment bv LC/MS/MS 12.0 Chromatography 12.1 Inject the same amount o f each standard, sample and fortified sample into the LC/MS/MS system. A calibration standard must precede and follow all analyzed samples. 12.2 Standards o f PFOA corresponding to at least five or more concentration levels must be included in an analytical set. 12.3 An entire set o f extracted calibration standards must be included at the beginning and at the end o f a sample set. Standards must be interspersed between every 5-10 samples. As an alternative, an entire set of calibration standards may be injected at the beginning o f a set followed by calibration standards interspersed every 5-10 samples (to account for a second set of standards). In either case, calibration standards must be the first and last injection in a sample set. 12.4 Use linear standard curves for quantitation. Linear standard curves are generated for the analyte by linear regression using 1/x weighting o f peak area versus calibration standard concentration using MassLynx 3.3 (or equivalent) software system. 12.5 Sample response should not exceed standard responses. Any samples that exceed standard responses should be further diluted and reanalyzed. 13.0 Acceptance Criteria 13.1 Chromatogram must show a peak o f a daughter ion at 369 amu from a parent of 413 amu. The 413 amu parent corresponds to the PFOA anion, while the daughter ion (369 amu) represents the loss o f carbon dioxide. 13.2 Method blanks must not contain PFOA at levels greater than the LOQ. If a blank contains PFOA at levels greater than 0.2 ng/mL, then a new blank sample must be obtained and the entire set must be re-extracted. 13.3 Recoveries o f control spikes and matrix spikes must be between 70-130% of their known values. If a control spike falls outside the acceptable limits, the entire set o f samples should be re-extracted. Any matrix spike outside 70 130% should be evaluated by the analyst to determine if re-extraction is warranted. 13.4 Any calibration standard found to be a statistical outlier by using the Huge Error Test, may be excluded from the calculation o f the calibration curve However, the total number o f extracted calibration standards that could be excluded must not exceed 20% o f the total number of extracted standards injected. 13.5 The correlation coefficient (R) for calibration curves generated must be >0.992 (R2 >0.985). If calibration results fall outside these limits, then appropriate steps must be taken to adjust instrument operation, and the standards or the relevant set of samples should be reanalyzed. Page 6 of 7 E05-0210 Interim Report#13 Tennessee River Page 35 o f 65 Page 126 of 160 3M Environm ental Laboratory E05-0210 Interim Report#13 Exygen Protocol Number: P 0001131 Exygen Research Method Number V0001782 ANALYTICAL METHOD Method o f Analysis for the Determination o f Perfluorooctanoic Acid (PFOA) in Sediment by LC /M S/M S 13.6 Retention times between standards and samples must not drift more than 4 % within an analytical run. If retention time drift exceeds this limit within an analytical run then the set must be reanalyzed. 14.0 Calculations 14.1 Use the following equation to calculate the amount o f PFOA found (in ng/mL, based on peak area) using the standard curve (linear regression parameters) generated by the Mass Lynx software program: PFOA found (ng/mL) = (Peak area - intercept) x DF slope DF = factor by which the final volume was diluted, if necessary. 14.2 For samples fortified with known amounts o f PFOA prior to extraction, use the following equation to calculate the percent recovery. Recovery (%) = [ total analyte found (ng/mL) - analyte found in control (ng/mL)] ^ ]QQ analyte added (ng/mL) 14.3 Use the following equation to convert the amount o f PFOA found in ng/mL to ng/g (ppb). PFOA found (ppb) = fPFOA found (ng/mL) x final volume (5 mL)l sample weight (5 g) 14.4 Use the following equation (if necessary) to calculate the amount of PFOA found in ppb based on dry weight. PFOA found (ppb) dry weight = PFOA found (ppb) x [100% / total solids(%)] Page 7 of 7 E05-0210 Interim Report#13 Tennessee River Page 36 o f 65 Page 127 of 160 3M Environm ental Laboratory E05-0210 Interim Report#13 Exygen Protocol Number: P0001131 ANALYTICAL METHOD Method Number: V0001783 Method of Analysis for the Determination of Perfluorooctanoic Acid (PFOA) in Fish and Clams by LC/MS/MS Analytical Testing Facility: Exygen Research 3058 Research Drive State College, PA 16801 Approved By: f Jl.. Paul Connolly ' Technical Leader, LC-MS, Exygen Research I Iz-fe/cui Date '/ m /d Y *ohn Flaherty Vice President, Operations, Exygen Research Date Total Pages: 8 E05-0210 Interim Report#13 Tennessee River Page 37 o f 65 Page 128 of 160 3M Environm ental Laboratory E05-0210 Interim Report#13 Exygen Protocol Number: P 0001131 Exygen Research Method Number V0001783 | ANALYTICAL m e t h o d Method o f Analysis for the Determination o f Perfluorooctanoic Acid (PFOA) in Fish and Clams by LC/MS/MS 1.0 Scope This method is to be employed for the isolation and quantitation o f perfluorooctanoic acid by High Performance Liquid Chromatography coupled to a tandem Mass Spectrometric Detector (LC/MS/MS) in fish and clams. 2.0 Safety 2.1 Always observe safe laboratory practices. 2.2 Consult the appropriate MSDS before handling any chemical for proper safety precautions. 3.0 Sample Requirement 3.1 At least 20 g o f test sample for extraction. 3.2 Samples should be processed before extraction. Place the frozen sample in a food processor and homogenize with dry ice. Place the samples in containers and leave open in frozen storage overnight to allow for carbon dioxide sublimation. Seal and place the samples in frozen storage until time of analysis. 3.3 Sample collection procedures will be specified in the sampling plan for this project. 4.0 Reagents and Standards 4.1 Water - HPLC grade 4.2 Acetonitrile - HPLC grade 4.3 Carbon (120-400 mesh) - Reagent grade 4.4 Methanol - HPLC grade 4.5 Silica gel (60-200 mesh) - Reagent grade 4.6 Florisil (60-100 mesh) - Reagent grade 4.7 Superclean LC-NH2 - Reagent grade 4.8 1-Octanol - HPLC grade 4.9 L-Ascorbic acid - Reagent grade 4.10 Dimethyldichlorosilane - Reagent grade 4.11 Toluene - Reagent grade 4.12 Ammonium Acetate - A.C.S. Reagent Grade 4.13 Perfluorooctanoic Acid - Sigma-Aldrich 5.0 Instrument and Equipment 5.1 A high performance liquid chromatograph capable of pumping up to 2 solvents equipped with a variable volume injector capable of injecting 5-200 pL connected to a tandem Mass Spectrometer (LC/MS/M S). 5.2 A device to collect raw data for peak integration and quantitation. 5.3 Analytical balance capable o f reading to 0.00001 g. Page 2 of S E05-0210 Interim Report#13 Tennessee River Page 38 o f 65 Page 129 of 160 3M Environm ental Laboratory E05-0210 Interim Report#13 Exygen Protocol Number: P0001131 Exygen Research Method NumberV0001783 1 ANALYTICAL METHOD Method of Analysis for the Determination o f Perfluorooctanoic Acid (PFOA) in Fish and Clams by LC/MS/MS 5.4 Rotary evaporator. 5.5 Tissumizer. 5.6 125 mL pear-shaped flasks. 5.7 50 mL disposable polypropylene centrifuge tubes. 5.8 15 mL disposable polypropylene centrifuge tubes. 5.9 Disposable micropipets (50-1 OOuL, 100-200uL). 5.10 125-mL LDPE narrow-mouth bottles. 5.11 2 mL clear HPLC vial kit. 5.12 Disposable pipettes. 5.13 Autopipettes (100-1000 pL and 10-100 pL), with disposable tips. 5.14 SPE tubes (20mL) (Supelco cat. no. N057177). 5.15 Wrist action shaker. 5.16 Centrifuge capable o f spinning 50 mL polypropylene tubes at 2000 rpm. 6.0 Chromatographic System 6.1 Analytical Column: Fluophase RP (Keystone Scientific), 2.1 mm x 50 mm, 5p (P/N: 82505-052130) 6.2 Temperature: 30C 6.3 Mobile Phase (A) : 2 mM Ammonium Acetate in Water 6.4 Mobile Phase (B) : Methanol 6.5 Gradient Program: Time (min) 0.0 1.0 8.0 20.0 22.5 %A 65 65 25 25 65 Flow Rate % B (mL/min) 35 0.3 35 0.3 75 0.3 75 0.3 35 0.3 6.6 Injection Volume: 15 pL (can be increased to as much as 50 pL). 6.7 Quantitation: Peak Area - external standard calibration curve. 6.8 Run Time: ~ 23 minutes. The above conditions are intended as a guide and may be changed in order to optimize the HPLC system. Page 3 of 8 E05-0210 Interim Report#13 Tennessee River Page 39 o f 65 Page 130 o f 160 3M Environm ental Laboratory E05-0210 Interim Report#13 Exygen Protocol Number: P0001131 Exygen Research Method Number VOOO1783 ANALYTICAL METHOD Method o f Analysis for the Determination o f Perfluorooctanoic Acid (PFOA) in Fish and Clams by LC/MS/MS 7.0 MS/MS System 7.1 Mode: Electrospray Negative MRM mode, monitoring 413 - 369 m/z for PFOA. The above conditions are intended as a guide and may be changed in order to optimize the MSMS system. 8.0 Preparation o f Solutions 8.1 Mobile Phase 8.1.1 2 mM ammonium acetate in water is prepared by adding 0.154 g of ammonium acetate to 1000 mL o f water. 8.2 Extraction Solutions 8.2.1 8.2.2 2% ascorbic acid in methanol is prepared by dissolving 2 g o f ascorbic acid in 100 mL o f methanol. 30% Dimethyldichlorosilane in toluene is prepared by bringing 3 mL o f dimethyldichlorosilane to a final volume o f 10 mL with toluene. Alternate volumes may be prepared. 9.0 Standard Preparation 9.1 Standard Stock/Fortification Solution 9.1.1 9.1.2 9.1.3 9.1.4 9.1.5 Prepare a stock solution o f -100 pg/mL o f PFOA by weighing 10 mg of analytical standard (corrected for purity) and dilute to 100 mL with methanol in a 125-mL LDPE bottle. A 1.0 pg/mL fortification solution o f PFOA is prepared by bnnging 1 mL o f the 100 pg/mL solution to a final volume o f 100 with m e th a n o l in a 125 mL LDPE bottle. A 0.1 pg/mL fortification solution o f PFOA is prepared by bringing 10 mL o f the 1.0 pg/mL solution to a final volume o f 100 with methanol in a 125 mL LDPE bottle. A 0.01 pg/mL fortification solution o f PFOA is prepared by bringing 10 mL o f the 0.1 pg/mL solution to a final volume of 100 with methanol in a 125 mL LDPE bottle. The stock and fortification solutions are to be stored in a refrigerator at approximately 4C and are stable for a maximum period of 6 months from the date o f preparation. Page 4 of 8 E05-0210 Interim Report#13 Tennessee River Page 40 o f 65 Page 131 of 160 3M Environm ental Laboratory E05-0210 Interim Report#13 Exygen Protocol Number: P0001131 Exygen Research Method Number V0001783 ANALYTICAL METHOD Method o f Analysis for the Determination o f Perfluorooctanoic Acid (PFOA) in Fish and Clams by LC/MS/MS 9.2 Standard Calibration Solutions 9.2.1 9.2.2 LC/MS/MS calibration standards are prepared in methanol via dilution of the 1.0 pg/mL fortification solution. The following is a typical example: additional concentrations may be prepared as needed. Concentration Final o f Fortification Volume Diluted to Concentration Solution (ug/mL) (mL) (mL) (pg/mL) 1.0 5.0 100 0.05 1.0 2.5 100 0.025 1.0 1.0 100 0.01 0.05 10 100 0.005 0.025 10 100 0.0025 0.1 10 100 0.001 0.005 10 100 0.0005 9.2.3 Store all calibration standards in 125-mL LDPE narrow-mouth bottles at 2C to 6C, up to six months. 9.2.4 Alternate volumes and concentrations o f standards may be prepared as needed. 10.0 Batch Set Up 10.1 Each batch o f samples extracted (typically 20 or less) must include at least one untreated control and two untreated controls fortified at known concentrations (lab control spike) to verify procedural recovery for the batch. 10.2 Requirements for field and laboratory duplicates and spikes will be specified in the quality assurance plan for this project. 11.0 Sample Extraction 11.1 Weigh 5 g of frozen sample into 50 mL polypropylene centrifuge tubes (fortify as needed, replace lid and mix well). 11.2 Add 30 mL o f acetonitrile and shake on a wrist action shaker for ~15 minutes. 11.3 Place the tubes in a freezer for ~1 hour. 11.4 Pack and condition the SPE tubes and silanize the pear-shaped flasks. 11.5 Pack the 20 mL SPE tubes in sequence with 2 g florisil, 2 g silica gel, 2 g carbon, and 1 g LC-NH2. Condition the columns with 20 mL of methanol, then 20 mL of acetonitrile. Discard all washes. Do not allow the column to dry. 11.6 Silanize the 125 mL pear-shaped flasks by rinsing with the 30% dimethyldichlorosilane in toluene solution. Rinse the flask with toluene once, followed by methanol (three times). Dry the flasks completely before use, either by air-drying or with a stream o f nitrogen. Page 5 of S E05-0210 Interim Report#13 Tennessee River Page 41 o f 65 P a g e 1 3 2 o f 1 6 0 .. 3M Environm ental Laboratory E05-0210 Interim Report#13 Exygen Protocol Number: P0001131 Exygen Research Method Number V0001783 ANALYTICAL METHOD ~ Method o f Analysis for the Determination o f Perfluorooctanoic Acid (PFOA) in Fish and Clams by LC/MS/MS 11.7 Centrifuge the 50 mL polypropylene tubes containing sample at -2000 rpm for ~10 minutes. 11.8 Decant the extract on to a conditioned SPE column fitted inside the mouth of the pear-shaped flask. Collect the eluate in the 125 mL silanized pear-shape flask. 11.9 Add 10 mL o f acetonitrile to the sample in the 50 mL centrifuge tube. Homogenize the frozen fat phase using a tissumizer for -3 0 seconds and rinse the tissumizer with -10 mL o f acetonitrile into the tube. 11.10 Shake the sample again for -10 minutes on a wrist-action shaker. 11.11 Place the tubes in a freezer for - 1 hour more. 11.12 Centrifuge the 50 mL polypropylene tubes containing sample at -2000 rpm fo r-10 minutes. 11.13 Decant the extract onto the same SPE column. Collect the eluate into the same pear-shaped flask and combine with the eluent from the initial extraction. 11.14 Pass 20 mL o f acetonitrile through the SPE column and combine the eluate in the same pear-shaped flask. 11.15 Add 3-4 drops o f 1-octanol to the extract in the pear-shaped flask and evaporate at reduced pressure using a rotary evaporator (at < 40C). 11.16 Make the final volume, by adding 2 mL o f 2% ascorbic acid in methanol to the pear-shaped flask and swirl to mix/dissolve. 11.17 Transfer the extracts to HPLC vials using disposable pipets. 11.18 Analyze samples using electrospray LC/M S/M S. 12.0 Chromatography 12.1 Inject the same amount o f each standard, sample and fortified sample into the LC/MS/MS system. A calibration standard must precede and follow all analyzed samples. 12.2 Standards o f PFOA corresponding to at least five or more concentration levels must be included in an analytical set. 12.3 An entire set o f calibration standards must be included at the beginning and at the end o f a sample set. Standards must be interspersed between every 5-10 samples. As an alternative, an entire set o f calibration standards may be injected at the beginning o f a set followed by calibration standards interspersed every 5-10 samples (to account for a second set o f standards). In either case, calibration standards must be the first and last injection in a sample set. 12.4 Use linear standard curves for quantitation. Linear standard curves are generated for the analyte by linear regression using 1/x weighting of peak area versus calibration standard concentration using MassLynx 3.3 (or equivalent) software system. Page 6 of 8 E05-0210 Interim Report#13 Tennessee River Page 42 o f 65 Page 133 of 160 3M Environm ental Laboratory E05-0210 Interim Report#13 Exygen Protocol Number: P0001131 Exygen Research Method Number V0001783 | ANALYTICAL METHOD Method o f Analysis for the Determination ofPerfluorooctanoic Acid (PFOA) in Fish and Clams by LC/MS/MS 12.5 Sample response should not exceed standard responses. Any samples that exceed standard responses should be further diluted and reanalyzed. 13.0 Acceptance Criteria 13.1 Chromatogram must show a peak o f a daughter ion at 369 amu from a parent o f 413 amu. The 413 amu parent corresponds to the PFOA anion, while the daughter ion (369 amu) represents the loss o f carbon dioxide. 13.2 Method blanks must not contain PFOA at levels greater than the LOQ. If a blank contains PFOA at levels greater than 0.5 ppb, then a new blank sample must be obtained and the entire set must be re-extracted. 13.3 Recoveries of control spikes and matrix spikes must be between 70-130% of their known values. If a control spike falls outside the acceptable limits, the entire set o f samples should be re-extracted. 13.4 Any calibration standard found to be a statistical outlier by using the Huge Error Test, may be excluded from the calculation o f the calibration curve However, the total number o f calibration standards that could be excluded must not exceed 20% o f the total number o f standards injected. 13.5 The correlation coefficient (R) for calibration curves generated must be >0.992 (R2 >0.985). If calibration results fall outside these limits, then appropriate steps must be taken to adjust instrument operation, and the standards or the relevant set o f samples should be reanalyzed. 13.6 Retention times between standards and samples must not drift more than 4 % within an analytical run. If retention time drift exceeds this limit within an analytical run then the set must be reanalyzed. 14.0 Calculations 14.1 Use the following equation to calculate the amount of PFOA found (in ng/mL, based on peak area) using the standard curve (linear regression parameters) generated by the Mass Lynx software program: PFOA found (ng/mL) = (Peak area - intercept) slope 14.2 Use the following equation to convert the amount o f PFOA found in ng/mL to ng/g (ppb). PFOA found (ppb) = [PFOA found (ng/mL) x final volume (mL) x DF1 sample weight (g) DF = factor by which the final volume was diluted, if necessary. Page 7 of 8 E 0 5 -0 2 1 0 In te rim 'R e p o rt# 1 3 Tennessee River Page 43 o f 65 Page 134 of 16 0 3M Environm ental Laboratory E05-0210 Interim Report#13 Exygen Protocol Number: P0001131 Exygen Research Method Number VOOO1783 1 a x a l y t i c a l .m e t h o d Method o f Analysis for the Determination o f Perfluorooctanoic Acid (PFOA) in Fish and Clams by LC/MS/MS 14.3 For samples fortified with known amounts o f PFOA prior to extraction, use the following equation to calculate the percent recovery. Recovery (%) = [ total analyte found (ng/g) - analyte found in control (ng/g)] ^ analyte added (ng/g) Page 8 of 8 E05-0210 Interim R eport#13 Tennessee River Page 44 o f 65 .... P a g e 1 3 5 o f 1 6 0 3M Environm ental Laboratory E05-0210 Interim Report#13 Exygen Protocol Number: P 0001131 ANALYTICAL METHOD Method Number: V0001784 Method of Analysis for the Determination of Perfluorooctanoic Acid (PFOA) in Vegetation by LC/MS/MS Analytical Testing Facility: Exygen Research 3058 Research Drive State College, PA 16801 Approved By: TLA CAL Pauilll PCnornmnonllllyv Technical Leader, LC-MS, Exygen Research n,//r>/fuC/ t>hn Flaherty Vice President, Operations, Exygen Research io)w >M Date Date Total Pages: 7 E05-0210 Interim Report#13 Tennessee River Page 45 o f 65 Page 136 of 160 3M Environm ental Laboratory E05-0210 Interim Report#13 Exygen Protocol Number: P 0001131 Exygen Research Method Number V0O01 784 1 ANALYTICAL METHOD Method o f Analysis for the Determination o f Perfluorooctanoic Acid (PFOA) in Vegetation by LC/MS/MS 1.0 Scope This method is to be employed for the isolation and quantitation o f perfluorooctanoic acid by High Performance Liquid Chromatography coupled to a tandem Mass Spectrometric Detector (LC/MS/MS) in vegetation. 2.0 Safety 2.1 Always observe safe laboratory practices. 2.2 Consult the appropriate M SDS before handling any chemical for proper safety precautions. 3.0 Sample Requirement 3.1 At least 20 g o f test sample for extraction. 3.2 Samples should be processed before extraction. Place the frozen sample in a food processor and homogenize with dry ice. Place the samples in containers and leave open in frozen storage overnight to allow for carbon dioxide sublimation. Seal and place the samples in frozen storage until time of analysis. 3.3 Sample collection procedures will be specified in the sampling plan for this project. 4.0 Reagents and Standards 4.1 Water - HPLC grade 4.2 Acetonitrile - HPLC grade 4.3 Carbon (120-400 mesh) - Reagent grade 4.4 Methanol - HPLC grade 4.5 Silica gel (60-200 mesh) - Reagent grade 4.6 Florisil (60-100 mesh) - Reagent grade 4.7 Superclean LC-NH2 - Reagent grade 4.8 1-Octanol - HPLC grade 4.9 L-Ascorbic acid - Reagent grade 4.10 Dimethyldichlorosilane - Reagent grade 4.11 To luene - Reagent grade 4.12 Ammonium Acetate - A.C.S. Reagent Grade 4.13 Perfluorooctanoic Acid - Sigma-Aldrich 5.0 Instrument and Equipment 5.1 A high performance liquid chromatograph capable o f pumping up to 2 solvents equipped with a variable volume injector capable o f injecting 5-200 pL connected to a tandem Mass Spectrometer (LC/MS/M S). 5.2 A device to collect raw data for peak integration and quantitation. 5.3 Analytical balance capable o f reading to 0.00001 g. Page 2 of 7 E05-0210 Interim Report#13 Tennessee River Page 46 o f 65 Page 137 of 160 3M Environm ental Laboratory E05-0210 Interim Report#13 Exygen Protocol Number: P 0001131 Exygen Research Method Number V0001784 ANALYTICAL METHOD Method o f Analysis for the Determination o f Perfluorooctanoic Acid (PFOA) in Vegetation by LC/MS/MS 5.4 Rotary evaporator. 5.5 125 mL pear-shaped flasks. 5.6 50 mL disposable polypropylene centrifuge tubes. 5.7 15 mL disposable polypropylene centrifuge tubes. 5.8 Disposable micropipets (50-1 OOuL, 100-200uL). 5.9 125-mLLDPE narrow-mouth bottles. 5.10 2 mL clear HPLC vial kit. 5.11 Disposable pipettes. 5.12 Autopipettes (100-1000 pL and 10-100 pL), with disposable tips. 5.13 SPE tubes (20mL) (Supelco cat. no. N057177). 5.14 Wrist action shaker. 5.15 Centrifuge capable o f spinning 50 mL polypropylene tubes at 2000 rpm. 6.0 Chromatographic System 6.1 Analytical Column: Fluophase RP (Keystone Scientific), 2.1 mm x 50 mm, 5p (P/N: 82505-052130) 6.2 Temperature: 30C 6.3 Mobile Phase (A) : 2 mM Ammonium Acetate in Water 6.4 Mobile Phase (B) : Methanol 6.5 Gradient Program: Time (min) 0.0 1.0 8.0 20.0 22.5 %A 65 65 25 25 65 Flow Rate % B (mL/min) 35 0.3 35 0.3 75 0.3 75 0.3 35 0.3 6.6 Injection Volume: 15 pL (can be increased to as much as 50 pL). 6.7 Quantitation: Peak Area - external standard calibration curve. 6.8 Run Time: - 23 minutes. The above conditions are intended as a guide and may be changed in order to optimize the HPLC system. 7.0 MS/MS System 7.1 Mode: Electrospray Negative MRM mode, monitoring 413 --> 369 m/z for PFOA. Page 3 of 7 E05-0210 Interim Report#13 Tennessee River Page 47 o f 65 P a g e 1 3 8 o f 1 6 0 . .. 3M Environm ental Laboratory E05-0210 Interim Report#13 Exygen Protocol Number: P 0001131 Exygen Research Method Number V0001784 | ANALYTICAL m e t h o d Method o f Analysis for the Determination o f Perfluorooctanoic Acid (PFOA) in Vegetation by LC/MS/MS The above conditions are intended as a guide and may be changed in order to optimize the MSMS system. 8.0 Preparation o f Solutions 8.1 Mobile Phase 8.1.1 2 mM ammonium acetate in water is prepared by adding 0.154 g of ammonium acetate to 1000 mL o f water. 8.2 Extraction Solutions 8.2.1 8.2.2 2% ascorbic acid in methanol is prepared by dissolving 2 g of ascorbic acid in 100 mL o f methanol. 30% Dimethyldichlorosilane in toluene is prepared by bringing 3 mL o f dimethyldichlorosilane to a final volume o f 10 mL with toluene. Alternate volumes may be prepared. 9.0 Standard Preparation 9.1 Standard Stock/Fortification Solution 9.1.1 9.1.2 9.1.3 9.1.4 9.1.5 Prepare a stock solution o f ~100 pg/mL o f PFOA by weighing 10 mg o f analytical standard (corrected for purity) and dilute to 100 mL with methanol in a 125-mL LDPE bottle. A 1.0 pg/mL fortification solution o f PFOA is prepared by bringing 1 mL o f the 100 pg/mL solution to a final volume o f 100 with methanol in a 125 mL LDPE bottle. A 0.1 pg/mL fortification solution o f PFOA is prepared by bringing 10 mL o f the 1.0 pg/mL solution to a final volume o f 100 with methanol in a 125 mL LDPE bottle. A 0.01 pg/mL fortification solution o f PFOA is prepared by bringing 10 mL of the 0.1 pg/mL solution to a final volume o f 100 with methanol in a 125 mL LDPE bottle. The stock and fortification solutions are to be stored in a refrigerator at approximately 4C and are stable for a maximum period o f 6 months from the date o f preparation. 9.2 Standard Calibration Solutions 9.2.1 LC/MS/MS calibration standards are prepared in methanol via dilution of the 1.0 pg/mL fortification solution. Page 4 ol E05-0210 Interim Report#13 Tennessee River Page 48 o f 65 Page 139 of 160 3M Environm ental Laboratory E05-0210 Interim Report#13 Exygen Protocol Number: P 0001131 Exygen Research Method Number V0001784 | A iNa l y h c 'a l ' m e t h o d Method o f Analysis for the Determination o f Perfluorooctanoic Acid (PFOA) in Vegetation by LC/MS/MS 9.2.2 The following is a typical example: additional concentrations may be prepared as needed. Concentration of Fortification Solution (ug/mL) 1.0 1.0 1.0 0.05 0.025 0.1 0.005 Volume (mL) 5.0 2.5 1.0 10 10 10 10 Diluted to (mL) 100 100 100 100 100 100 100 Final Concentration (pg/mL) 0.05 0.025 0.01 0.005 0.0025 0.001 0.0005 9.2.3 Store all calibration standards in 125-mL LDPE narrow-mouth bottles at 2C to 6C, up to six months. 9.2.4 Alternate volumes and concentrations o f standards may be prepared as needed. 10.0 Batch Set Up 10.1 Each batch o f samples extracted (typically 20 or less) must include at least one untreated control and two untreated controls fortified at known concentrations (lab control spike) to verify procedural recovery for the batch 10.2 Requirements for field and laboratory duplicates and spikes will be specified in the quality assurance plan for this project. 11.0 Sample Extraction 11.1 Weigh 5 g o f frozen sample into 50 mL polypropylene centrifuge tubes (fortify as needed, replace lid and mix well). 11.2 Add 30 mL of acetonitrile and shake on a wrist action shaker for -15 minutes. 11.3 Centrifuge the 50 mL polypropylene tubes containing sample at -2000 rpm fo r-1 0 minutes. 11.4 Pack and condition the SPE tubes and silanize the pear-shaped flasks. 11.5 Pack the 20 mL SPE tubes in sequence with 2 g florisil, 2 g silica gel, 2 g carbon, and 1 g LC-NH2. Condition the columns with 20 mL of methanol, then 20 mL o f acetonitrile. Discard all washes. Do not allow the column to dry. 11.6 Silanize the 125 mL pear-shaped flasks by rinsing with the 30/) dimethyldichlorosilane in toluene solution. Rinse the flask with toluene once, followed by methanol (three times). Dry the flasks completely before use, either by air-drying or with a stream o f nitrogen. 11.7 Decant the extract on to a conditioned SPE column fitted inside the mouth 01 ' the pear-shaped flask. Collect the eluate in the 125 mL silanized pear-shape flask. Page 5 o f " E05-0210 Interim Report#13 Tennessee River Page 49 o f 65 Page 140 of 160 3M Environm ental Laboratory E05-0210 Interim Report#13 Exygen Protocol Number: P 0001131 Exygen Research Method Number V0001?84 | ANALYTICAL ViF t HOD Method of Analysis for the Determination o f Perfluorooctanoic Acid (PFOA) in Vegetation by LC/MS/MS 11.8 Add 20 mL o f acetonitrile to the sample in the 50 mL centrifuge tube. 11.9 Shake the sample again for ~10 minutes on a wrist-action shaker. 11.10 Centrifuge the 50 mL polypropylene tubes containing sample at -2000 rpm for ~5 minutes. 11.11 Decant the extract onto the same SPE column. Collect the eluate into the same pear-shaped flask and combine with the eluent from the initial extraction. 11.12 Repeat steps 11.8 through 11.11 again. 11.13 Add 3-4 drops o f 1-octanol to the extract in the pear-shaped flask and evaporate at reduced pressure using a rotary evaporator (at < 40C). 11.14 Make the final volume, by adding 2 mL o f 2% ascorbic acid in methanol to the pear-shaped flask and swirl to mix/dissolve. 11.15 Transfer the extracts to HPLC vials using disposable pipets. 11.16 Analyze samples using electrospray LC/MS/MS. 12.0 Chromatography 12.1 Inject the same amount o f each standard, sample and fortified sample into the LC/MS/MS system. A calibration standard must precede and follow all analyzed samples. 12.2 Standards of PFOA corresponding to at least five or more concentration levels must be included in an analytical set. 12.3 An entire set o f extracted calibration standards must be included at the beginning and at the end o f a sample set. Extracted standards must be interspersed between every 5-10 samples. As an alternative, an entire set of extracted calibration standards may be injected at the beginning of a set followed by extracted calibration standards interspersed every 5-10 samples (to account for a second set o f extracted standards). In either case, extracted calibration standards must be the first and last injection in a sample set. 12.4 Use linear standard curves for quantitation. Linear standard curves are generated for the analyte by linear regression using 1/x weighting of peak area versus calibration standard concentration using MassLynx 3.3 (or equivalent) software system. 12.5 Sample response should not exceed standard responses. Any samples that exceed standard responses should be further diluted and reanalyzed. 13.0 Acceptance Criteria 13.1 Chromatogram must show a peak o f a daughter ion at 369 amu from a parent o f 413 amu. The 413 amu parent corresponds to the PFOA anion, while the daughter ion (369 amu) represents the loss o f carbon dioxide. Page 6 of 7 E05-0210 Interim Report#13 Tennessee River Page 50 o f 65 Page 141 of 160 3M Environm ental Laboratory E05-0210 Interim Report#13 Exygen Protocol Number: P0001 131 Exygen Research Method Number V0001784 I ANALYTICAL m e t h o d Method o f Analysis for the Determination o f Perfluorooctanoic Acid (PFOA) in Vegetation by LC/MS/MS 13.2 Method blanks must not contain PFOA at levels greater than the LOQ. If a blank contains PFOA at levels greater than 0.5 ppb, then a new blank sample must be obtained and the entire set must be re-extracted. 13.3 Recoveries o f control spikes and matrix spikes must be between 70-130% of their known values. If a control spike falls outside the acceptable limits, the entire set o f samples should be re-extracted. 13.4 Any calibration standard found to be a statistical outlier by using the Huge Error Test, may be excluded from the calculation of the calibration curve. However, the total number o f calibration standards that could be excluded must not exceed 20% of the total number o f standards injected. 13.5 The correlation coefficient (R) for calibration curves generated must be >0.992 (R2 >0.985). If calibration results fall outside these limits, then appropriate steps must be taken to adjust instrument operation, and the standards or the relevant set o f samples should be reanalyzed. 13.6 Retention times between standards and samples must not drift more than 4 % within an analytical run. If retention time drift exceeds this limit within an analytical run then the set must be reanalyzed. 14.0 Calculations 14.1 Use the following equation to calculate the amount o f PFOA found (in ng/mL, based on peak area) using the standard curve (linear regression parameters) generated by the Mass Lynx software program: PFOA found (ng/mL) = (Peak area - intercept) slope 14.2 Use the following equation to convert the amount o f PFOA found in ng/mL to ng/g (ppb). PFOA found (ppb) = rPFOA found (ng/mL) x final volume (mL) x DF1 sample weight (g) DF = factor by which the final volume was diluted, if necessary. 14.3 For samples fortified with known amounts o f PFOA prior to extraction, use the following equation to calculate the percent recovery. Recovery (%) = [ total analyte found (ng/g) - analyte found in control (ng/g)] QQ analyte added (ng/g) Page 7 oi' 7 E05-0210 Interim Report#13 Tennessee River Page 51 o f 65 Page 142 of 160 3M Environm ental Laboratory E05-0210 Interim Report#13 Exygen Protocol Number: P00113 1 ANALYTICAL METHOD Method Number: V0001785 Method of Analysis for the Determination of Perfluorooctanoic Acid (PFOA) in Small Mammal Liver by LC/MS/MS Analytical Testing Facility: Exygen Research 3058 Research Drive State College, PA 16801 Approved By: Paul Connolly Technical Leader, LC-MS, Exygen Research a// f i/Z / o 'm . Flaherty ' Vice President, Operations, Exygen Research Date Date Total Pages: 7 E05-0210 Interim Report#13 Tennessee River Page 52 o f 65 .P a g e 1 4 3 o f 1 6 0 3M Environm ental Laboratory E05-0210 Interim Report#13 Exygen Protocol Number: P 0001131 Exygen Research Method Number V0001785 A^LYTICAL^M ETHO D Method o f Analysis for the Determination o f Perfluorooctanoic Acid (PFOA) in Small Mammal Liver by LC/MS/MS 1.0 Scope This method is to be employed for the isolation and quantitation o f perfluorooctanoic acid by High Performance Liquid Chromatography coupled to a tandem Mass Spectrometric Detector (LC/MS/MS) in small mammal liver. 2.0 Safety 2.1 Always observe safe laboratory practices. 2.2 Consult the appropriate M SDS before handling any chemical for proper safety precautions. 3.0 Sample Requirement 3.1 At least 5 g o f test sample for extraction. 3.2 Samples should be processed before extraction. Place the frozen sample in a food processor and homogenize with dry ice. Place the samples in containers and leave open in frozen storage overnight to allow for carbon dioxide sublimation. Seal and place the samples in frozen storage until time of analysis. Alternately, if there is an insufficient amount o f sample (-less than 5 g), then no processing is necessary and the sample can be used as supplied. 3.3 Sample collection procedures will be specified in the sampling plan for this project. 4.0 Reagents and Standards 4.1 Water - HPLC grade 4.2 Methanol - HPLC grade 4.3 Acetonitrile - HPLC grade 4.4 Ammonium Acetate - A.C.S. Reagent Grade 4.5 Perfluorooctanoic Acid - Sigma-Aldrich 5.0 Instrument and Equipment 5.1 A high performance liquid chromatograph capable o f pumping up to 2 solvents equipped with a variable volume injector capable of injecting 5-200 pL connected to a tandem Mass Spectrometer (LC/MS/M S). 5.2 A device to collect raw data for peak integration and quantitation. 5.3 Analytical balance capable of reading to 0.00001 g. 5.4 50 mL disposable polypropylene centrifuge tubes. 5.5 15 mL disposable polypropylene centrifuge tubes. 5.6 Disposable micropipets (50-1 OOuL, 100-200uL). 5.7 125-mLLDPE narrow-mouth bottles. 5.8 2 mL clear HPLC vial kit. Page 2 of"7 E05-0210 Interim Report#13 Tennessee River Page 53 o f 65 Page 144 of 160 3M Environm ental Laboratory E05-0210 Interim Report#13 Exygen Protocol Number: P0001131 Exygen Research Method Number V0001785 | ANAi7v T IC ^ l "m THQD Method o f Analysis for the Determination o f Perfluorooctanoic Acid (PFOA) in Small Mammal Liver by LC/MS/MS 5.9 Disposable pipettes. 5.10 Autopipettes (100-1000 pL and 10-100 pL), with disposable tips. 5.11 Waters Sep Pak Vac 6 cc (lg ) tC18 SPE cartridges. 5.12 SPE vacuum manifold. 5.13 Tissuemizer. 5.14 Wrist-action shaker. 5.15 Centrifuge capable of spinning 15 mL polypropylene tubes at 3000 rpm. 6.0 Chromatographic System 6.1 Analytical Column: Fluophase RP (Keystone Scientific), 2.1 mm x 50 mm, 5p (P/N: 82505-052130) 6.2 Temperature: 30C 6.3 Mobile Phase ( A ) : 2 mM Ammonium Acetate in Water 6.4 Mobile Phase ( B ) : Methanol 6.5 Gradient Program: Time (min) 0.0 1.0 8.0 20.0 22.5 %A 65 65 25 25 65 Flow Rate % B (mL/min) 35 0.3 35 0.3 75 0.3 75 0.3 35 0.3 6.6 Injection Volume: 15 pL (can be increased to as much as 50 pL). 6.7 Quantitation: Peak Area - external standard calibration curve. 6.8 Run Time: ~ 23 minutes. The above conditions are intended as a guide and may be changed in order to optimize the HPLC system. 7.0 MS/MS System 7.1 Mode: Electrospray Negative MRM mode, monitoring 413 --^ 369 m/z for PFOA. The above conditions are intended as a guide and may be changed in order to optimize the MSMS system. Page 3 of 7 E05-0210 Interim Report#13 Tennessee River Page 54 u f 65 Page 145 of 160 3M Environm ental Laboratory E05-0210 Interim Report#13 Exygen Protocol Number: P 0001131 Exygen Research Method Number V0001785 1 AN vLYT1CAl "m T H QD Method of Analysis for the Determination ofPerfluorooctanoic Acid (PFOA) in Small Mammal Liver by LC/MS/MS 8.0 Preparation o f Solutions 8.1 Mobile Phase 8.1.1 2 mM ammonium acetate in water is prepared by adding 0.154 g of ammonium acetate to 1000 mL o f water. Alternate volumes may be prepared. 9.0 Standard Preparation 9.1 Standard Stock/Fortification Solution 9.1.1 Prepare a stock solution of -100 pg/mL o f PFOA by weighing 10 mg of analytical standard (corrected for purity) and dilute to 100 mL with methanol in a 125-mL LDPE bottle. 9.1.2 A 1.0 pg/mL fortification solution of PFOA is prepared by bringing 1 mL of the 100 pg/mL solution to a final volume o f 100 with methanol in a 125 mL LDPE bottle. 9.1.3 A 0.1 pg/mL fortification solution o f PFOA is prepared by bringing 10 mL o f the 1.0 pg/mL solution to a final volume o f 100 with methanol in a 125 mL LDPE bottle. 9.1.4 The stock and fortification solutions are to be stored in a refrigerator at approximately 4C and are stable for a maximum period o f 6 months from the date o f preparation. 9.2 Standard Calibration Solutions 9.2.1 9.2.2 LC/MS/MS calibration standards are prepared in methanol via dilution of the 0.1 pg/mL fortification solution. The following is a typical example: additional concentrations may be prepared as needed. Concentration of Fortification Solution (ng/mL) 100 100 100 5.0 2.0 1.0 Volume (mL) 5.0 2.0 1.0 10 10 10 Diluted to (mL) 100 100 100 100 100 100 Final Concentration (ng/mL) 5.0 2.0 1.0 0.5 0.2 0.1 9.2.3 Store all calibration standards in 125-mL LDPE narrow-mouth bottles at 2C to 6C, up to six months. 9.2.4 Alternate volumes and concentrations o f standards may be prepared as needed. Page 4 of 7 E05-0210 Interim Report#13 Tennessee River Page 55 o f 65 Page 146 of 160 3M Environm ental Laboratory E05-0210 Interim Report#13 Exygen Protocol Number: P 0001131 Exygen Research Method Number V0001785 | ANALYTICAL METHOD Method o f Analysis for the Determination ofPerfluorooctanoic Acid (PFOA) in Small Mammal Liver by LC/MS/MS j 10.0 Batch Set Up 10.1 Each batch o f samples extracted (typically 20 or less) must include at least one untreated control and two untreated controls fortified at known concentrations (lab control spike) to verify procedural recovery for the batch. 10.2 Requirements for field and laboratory duplicates and spikes will be specified in the quality assurance plan for this project. 11.0 Sample Extraction 11.1 Weigh 1 g of sample into a 50 mL polypropylene centrifuge tubes (fortify as needed, replace lid and mix well). Note that alternate weights of liver may be measured depending on the sample size available for use. 11.2 Add water to the sample for a final volume o f 10 mL. 11.3 Homogenize sample using a tissuemizer for ~ 1 minute. 11.4 Transfer 1 mL o f the sample using a disposable pipette into a 15 mL disposable centrifuge tube. 11.5 Add 5 mL o f acetonitrile and shake for ~20 minutes on a wrist-action shaker. 11.6 Centrifuge the tubes at -3000 rpm for -5 minutes. 11.7 Decant the supernatant into a 50 mL disposable centrifuge tube and add 35 mL o f water. 11.8 Condition the C 18 SPE cartridges (1 g, 6 mL) by passing 10 mL methanol followed by 5 mL o f HPLC water (~ 2 drop/sec). Do not let column run dry 11.9 Load the sample on conditioned Cig SPE cartridge. Discard eluate. 11.10 Elute with -2 mL of methanol. Collect 2 mL o f eluate into a graduated 15 mL polypropylene centrifuge tube (final volume = 2 mL). 11.11 Analyze samples using electrospray LC/MS/MS. 12.0 Chromatography 12.1 Inject the same amount o f each standard, sample and fortified sample into the LC/MS/MS system. A calibration standard must precede and follow all analyzed samples. 12.2 Standards o f PFOA corresponding to at least five or more concentration lev els must be included in an analytical set. 12.3 An entire set o f calibration standards must be included at the beginning and at the end of a sample set. Standards must be interspersed between every 5-10 samples. As an alternative, an entire set of calibration standards may be injected at the beginning o f a set followed by calibration standards interspersed every 5-10 samples (to account for a second set o f standards). In either case, calibration standards must be the first and last injection in a sample set. 12.4 Use linear standard curves for quantitation. Linear standard curves are generated for the analyte by linear regression using 1/x weighting of peak area Page 5 of 7 E05-0210 Interim Report#13 Tennessee River Page 56 o f 65 Page 147 of 160. 3M Environm ental Laboratory E05-0210 Interim Report#13 Exygen Protocol Number: P0001131 Exygen Research Method Number V0001785 | ANALYTICAL METHOD Method o f Analysis for the Determination ofPerfluorooctanoic Acid (PFOA) in Small Mammal Liver by LC/M S/M S versus calibration standard concentration using MassLynx 3.3 (or equivalent) software system. 12.5 Sample response should not exceed standard responses. Any samples that exceed standard responses should be further diluted and reanalyzed. 13.0 Acceptance Criteria 13.1 Chromatogram must show a peak o f a daughter ion at 369 amu from a parent o f 413 amu. The 413 amu parent corresponds to the PFOA anion, while the daughter ion (369 amu) represents the loss o f carbon dioxide. 13.2 Method blanks must not contain PFOA at levels greater than the LOQ. If a blank contains PFOA at levels greater than 10 ng/g, then a new blank sample must be obtained and the entire set must be re-extracted. 13.3 Recoveries o f control spikes and matrix spikes must be between 70-130% of their known values. If a control spike falls outside the acceptable limits, the entire set o f samples should be re-extracted. Any matrix spike outside 70 130% should be evaluated by the analyst to determine if re-extraction is warranted. 13.4 Any calibration standard found to be a statistical outlier by using the Huge Error Test, may be excluded from the calculation o f the calibration curve. However, the total number o f calibration standards that could be excluded must not exceed 20% o f the total number o f standards injected. 13.5 The correlation coefficient (R) for calibration curves generated must be >0.992 (R2 >0.985). If calibration results fall outside these limits, then appropriate steps must be taken to adjust instrument operation, and the standards or the relevant set o f samples should be reanalyzed. 13.6 Retention times between standards and samples must not drift more than 4 % within an analytical run. If retention time drift exceeds this limit within an analytical run then the set must be reanalyzed. 14.0 Calculations 14.1 Use the following equation to calculate the amount of PFOA found (in ng/rnL, based on peak area) using the standard curve (linear regression parameters) generated by the Mass Lynx software program: PFOA found (ng/mL) = fPeak area - intercept) x DF x aliquot factor slope DF = factor by which the final volume was diluted, if necessary. Aliquot factor = 10 Page 6 of 7 E05-0210 Interim Report#13 Tennessee River Page 57 o f 65 P a g e 1 4 8 . f i r n 3M Environm ental Laboratory E05-0210 Interim Report#13 Exygen Protocol Number: P 0001131 Exygen Research Method Number V0001785 | ANALY i 11AL METHOD Method of Analysis for the Determination o f Perfluorooctanoic Acid (PFOA) in Small Mammal Liver by LC/M S/M S j 14.2 For samples fortified with known amounts o f PFOA prior to extraction, use the following equation to calculate the percent recovery. Recovery (%) = [ total analyte found (ng/mL) - analyte found in control (ng/mL)] analyte added (ng/mL) 14.3 Use the following equation to convert the amount o f PFOA found in ng/mL to ng/g (ppb). PFOA found (ppb) = [PFOA found (ng/mL) x final volume (mL)l sample weight (g) Page 7 of 7 E05-0210 Interim R eport#13 Tennessee River Page 58 o f 65 Page 149 o f '160 3M Environm ental Laboratory E05-0210 Interim Report#13 Exygen Protocol Number: P 0001131 ANALYTICAL METHOD Method Number: V0001786 Method of Analysis for the Determination of Perfluorooctanoic Acid (PFOA) in Sm all Mammal Serum by LC/MS/MS Analytical Testing Facility: Exygen Research 3058 Research Drive State College, PA 16801 Approved By: ^ --V Paul Connolly Technical Leader, LC-MS, Exygen Research J /V / f / Z John Flaherty ' Vice President, Operations, Exygen Research \ q |z>(cn{ Date Date Total Pages: 7 E05-0210 Interim Report#13 Tennessee River Page 59 o f 65 Page 150 of 160 3M Environm ental Laboratory E05-0210 Interim Report#13 Exygen Protocol Number: P 0001131 Exygen Research Method Number V0001786 | ANALYTICAL METHOD Method o f Analysis for the Determination o f Perfluorooctanoic Acid (PFOA) in Small Mammal Serum by LC/M S/M S 1.0 Scope This method is to be employed for the isolation and quantitation of perfluorooctanoic acid by High Performance Liquid Chromatography coupled to a tandem Mass Spectrometric Detector (LC/MS/M S) in small mammal serum. 2.0 Safety 2.1 Always observe safe laboratory practices. 2.2 Consult the appropriate M SDS before handling any chemical for proper safety precautions. 3.0 Sample Requirement 3.1 At least 1 mL o f test sample for extraction. 3.2 No sample processing is needed for serum samples. However, frozen serum samples must to allowed to completely thaw to room temperature before use. 3.3 Sample collection procedures will be specified in the sampling plan for this project. 4.0 Reagents and Standards 4.1 Water - HPLC grade 4.2 Methanol - HPLC grade 4.3 Acetonitrile - HPLC grade 4.4 Ammonium Acetate - A.C.S. Reagent Grade 4.5 Perfluorooctanoic Acid - Sigma-Aldrich 5.0 Instrument and Equipment 5.1 A high performance liquid chromatograph capable o f pumping up to 2 solvents equipped with a variable volume injector capable of injecting 5-200 pL connected to a tandem Mass Spectrometer (LC/MS/M S). 5.2 A device to collect raw data for peak integration and quantitation. 5.3 Analytical balance capable o f reading to 0.00001 g. 5.4 50 mL disposable polypropylene centrifuge tubes. 5.5 15 mL disposable polypropylene centrifuge tubes. 5.6 Disposable micropipets (50-1 OOuL, 100-200uL). 5.7 125-mLLDPE narrow-mouth bottles. 5.8 2 mL clear HPLC vial kit. 5.9 Disposable pipettes. 5.10 Autopipettes (100-1000 pL and 10-100 pL), with disposable tips. 5.11 Waters Sep Pak Vac 6 cc (lg ) tC18 SPE cartridges. 5.12 SPE vacuum manifold. 5.13 Vortexer. Page 2 of 7 E05-0210 Interim Report#13 Tennessee River Page 60 o f 65 ...... 151.......... o f 160 3M Environm ental Laboratory E05-0210 Interim Report#13 Exygen Protocol Number: P 0001131 Exygen Research Method Number V0001786 | AiN/vlYTlCALM ETHOD Method o f Analysis for the Determination o f Perfluorooctanoic Acid (PFOA) in Small Mammal Serum by LC/MS/MS 5.14 Wrist-action shaker. 5.15 Centrifuge capable o f spinning 15 mL polypropylene tubes at 3000 rpm. 6.0 Chromatographic System 6.1 Analytical Column: Fluophase RP (Keystone Scientific), 2.1 mm x 50 mm, 5p (P/N: 82505-052130) 6.2 Temperature: 30C 6.3 Mobile Phase ( A ) : 2 mM Ammonium Acetate in Water 6.4 Mobile Phase ( B ) : Methanol 6.5 Gradient Program: Time (min) 0.0 1.0 8.0 20.0 22.5 %A 65 65 25 25 65 Flow Rate % B (mL/min) 35 0.3 35 0.3 75 0.3 75 0.3 35 0.3 6.6 Injection Volume: 15 pL (can be increased to as much as 50 pL). 6.7 Quantitation: Peak Area - external standard calibration curve. 6.8 Run Time: ~ 23 minutes. The above conditions are intended as a guide and may be changed in order to optimize the HPLC system. 7.0 MS/MS System 7.1 Mode: Electrospray Negative MRM mode, monitoring 413 -> 369 mIz for PFOA. The above conditions are intended as a guide and may be changed in order to optimize the MSMS system. 8.0 Preparation o f Solutions 8.1 Mobile Phase 8.1.1 2 mM ammonium acetate in water is prepared by adding 0.154 g of ammonium acetate to 1000 mL o f water. Alternate volumes may be prepared. Page 3 of 7 E05-0210 Interim Report#13 Tennessee River Page 61 o f 65 Page 152 of 160 3M Environm ental Laboratory E05-0210 Interim Report#13 Exygen Protocol Number: P 0001131 Exygen Research Method Number V0001786 ANALYTICAL METHOD Method o f Analysis for the Determination o f Perfluorooctanoic Acid (PFOA) in Small Mammal Serum by LC/M S/M S 9.0 Standard Preparation 9.1 Standard Stock/Fortification Solution 9.1.1 Prepare a stock solution o f -100 pg/mL o f PFOA by weighing 10 mg of analytical standard (corrected for purity) and dilute to 100 mL with methanol in a 125-mL LDPE bottle. 9.1.2 A 1.0 pg/mL fortification solution o f PFOA is prepared by bringing 1 mL of the 100 pg/mL solution to a final volume of 100 with methanol in a 125 mL LDPE bottle. 9.1.3 A 0.1 pg/mL fortification solution o f PFOA is prepared by bringing 10 mL o f the 1.0 pg/mL solution to a final volume o f 100 with methanol in a 125 mL LDPE bottle. 9.1.4 The stock and fortification solutions are to be stored in a refrigerator at approximately 4C and are stable for a maximum period of 6 months from the date o f preparation. 9.2 Standard Calibration Solutions 9.2.1 9.2.2 LC/MS/MS calibration standards are prepared in methanol via dilution of the 0.1 pg/mL fortification solution. The following is a typical example: additional concentrations may be prepared as needed. Concentration o f Fortification Solution (ng/mL) Volume (mL) Diluted to (mL) Final Concentration (ng/mL) 100 5.0 100 100 2.0 100 100 1.0 100 5.0 10 100 2.0 10 100 1.0 10 100 5.0 2.0 1.0 0.5 0.2 0.1 9.2.3 Store all calibration standards in 125-mL LDPE narrow-mouth bottles at 2C to 6C, up to six months. 9.2.4 Alternate volumes and concentrations o f standards may be prepared as needed. 10.0 Batch Set Up 10.1 Each batch o f samples extracted (typically 20 or less) must include at least one untreated control and two untreated controls fortified at known concentrations (lab control spike) to verify procedural recovery for the batch 10.2 Requirements for field and laboratory duplicates and spikes will be specified in the quality assurance plan for this project. Page 4 of 7 E05-0210 Interim Report#13 Tennessee River Page 62 o f 65 P a g e 1 5 3 e f i 60 3M Environm ental Laboratory E05-0210 Interim Report#13 Exygen Protocol Number: P 0001131 Exygen Research Method Number VOOO1786 I ANALYTICAL m e t h o d Method o f Analysis for the Determination o f Perfluorooctanoic Acid (PFOA) in Small Mammal Serum by LC/M S/M S 11.0 Sample Extraction 11.1 Measure 1 mL o f sample into a 50 mL polypropylene centrifuge tubes (fortify as needed, replace lid and mix well). Note that alternate volumes of serum may be measured depending on the sample size available for use. 11.2 Add water to the sample for a final volume o f 20 mL. Cap tightly 11.3 Vortex for ~1 minute. 11.4 Transfer 1 mL o f the sample using a disposable pipette into a 15 mL disposable centrifuge tube. 11.5 Add 5 mL o f acetonitrile and shake for ~20 minutes on a wrist-action shaker. 11.6 Centrifuge the tubes at -3000 rpm for -5 minutes. 11.7 Decant the supernatant into a 50 mL disposable centrifuge tube and add 35 mL o f water. 11.8 Condition the Cig SPE cartridges (1 g, 6 mL) by passing 10 mL methanol followed by 5 mL o f HPLC water (~ 2 drop/sec). Do not let column run dry 11.9 Load the sample on conditioned Cis SPE cartridge. Discard eluate. 11.10 Elute with -2 mL o f methanol. Collect 2 mL o f eluate into a graduated 15 mL polypropylene centrifuge tube (final volume = 2 mL). 11.11 Analyze samples using electrospray LC/MS/MS. 12.0 Chromatography 12.1 Inject the same amount o f each standard, sample and fortified sample into the LC/MS/MS system. A calibration standard must precede and follow all analyzed samples. 12.2 Standards o f PFOA corresponding to at least five or more concentration levels must be included in an analytical set. 12.3 An entire set o f calibration standards must be included at the beginning and at the end of a sample set. Standards must be interspersed between every 5-10 samples. As an alternative, an entire set of calibration standards may be injected at the beginning o f a set followed by calibration standards interspersed every 5-10 samples (to account for a second set of standards). In either case, calibration standards must be the first and last injection in a sample set. 12.4 Use linear standard curves for quantitation. Linear standard curves are generated for the analyte by linear regression using 1/x weighting of peak area versus calibration standard concentration using MassLynx 3.3 (or equivalent) software system. 12.5 Sample response should not exceed standard responses. Any samples that exceed standard responses should be further diluted and reanalyzed. Page 5 of 7 E 0 5 _0 2 io iM e r 1 m R e p o rt# 1 3 Tennessee River Page 63 o f 65 P a g e ig z o rrm 3M Environm ental Laboratory E05-0210 Interim Report#13 Exygen Protocol Number: P 0001131 Exygen Research Method Number V0001786 | ANAUmCALMKTHOD Method o f Analysis for the Determination o f Perfluorooctanoic Acid (PFOA) in Small Mammal Serum by LC/MS/MS 13.0 Acceptance Criteria 13.1 Chromatogram must show a peak o f a daughter ion at 369 amu from a parent o f 413 amu. The 413 amu parent corresponds to the PFOA anion, while the daughter ion (369 amu) represents the loss of carbon dioxide. 13.2 Method blanks must not contain PFOA at levels greater than the LOQ. If a blank contains PFOA at levels greater than 10 ng/mL, then a new blank sample must be obtained and the entire set must be re-extracted. 13.3 Recoveries o f control spikes and matrix spikes must be between 70-130% of their known values. If a control spike falls outside the acceptable limits, the entire set o f samples should be re-extracted. Any matrix spike outside 70 130% should be evaluated by the analyst to determine if re-extraction is warranted. 13.4 Any calibration standard found to be a statistical outlier by using the Huge Error Test, may be excluded from the calculation o f the calibration curve However, the total number o f calibration standards that could be excluded must not exceed 20% o f the total number o f standards injected. 13.5 The correlation coefficient (R) for calibration curves generated must be >0.992 (R2 >0.985). If calibration results fall outside these limits, then appropriate steps must be taken to adjust instrument operation, and the standards or the relevant set of samples should be reanalyzed. 13.6 Retention times between standards and samples must not drift more than 4 % within an analytical run. If retention time drift exceeds this limit within an analytical run then the set must be reanalyzed. 14.0 Calculations 14.1 Use the following equation to calculate the amount o f PFOA found (in ng/mL, based on peak area) using the standard curve (linear regression parameters) generated by the Mass Lynx software program: PFOA found (ng/mL) = (Peak area - intercept) x DF x aliquot factor slope DF = factor by which the final volume was diluted, if necessary. Aliquot factor = 20 14.2 For samples fortified with known amounts o f PFOA prior to extraction, use the following equation to calculate the percent recovery. Recovery (%) = [ total analyte found (ng/mL) - analyte found in control (ng/mL)] QQ analyte added (ng/mL)_____________________ _________ -- Page 6 of 7 E05-0210 Interim Report#13 Tennessee River Page 64 o f 65 Page 155 of 160 3M Environm ental Laboratory E05-0210 Interim Report#13 Exygen Protocol Number: P 0001131 Exygen Research Method Number V0001786 I ANa LVTICAL METHOD ~ Method o f Analysis for the Determination o f Perfluorooctanoic Acid (PFOA) in Small Mammal Serum by LC/MS/MS 14.3 Use the following equation to convert the amount o f PFOA found in ng/mL to ppb. PFOA found (ppb) = IPFOA found (ng/mL) x final volume (mL)1 sample volume (mL) Page 7 o f' E05-0210 Interim Report#13 Tennessee River Page 65 o f 65 Page 156 of 160 3M Environm ental Laboratory E05-0210 Interim R eport#13 Protocol Exygen P0001131; 3M Study Number E05-0210 Amendment 3 Study Title An a l y s is o f Pe r f l u o r o b u t a n e s u l f o n a t e (PFBS), Pe r f l u o r o h e x a n e s u l f o n a t e (PFHS), a n d PERFLUOROOCTANESULFONATE (PFOS) IN WATER, SO IL, SEDIMENT, FISH, CLAMS, Ve g e t a t io n , Sm a l l Ma m m a l Liv e r a n d Sm a l l Ma m m a l Se r u m Us in g LC/MS/MS f o r t h e 3M D M Pe c a t u r o n it o r in g r o g r a m PROTOCOL AMENDMENT NO. 3 Amendment Date: May 16, 2005 Performing Laboratory 3M Environmental, Health, and Safety Operations 3M Environmental Laboratory 935 Bush Avenue St. Paul, MN 55106 Laboratory Project Identification E05-0210 E05-0210 E05-0210 Interim R eport#13 GLP Protocol Amendment #3 Tennessee R iver Page 1 of 4 Page 157 of 160 3M Environm ental Laboratory E05-0210 Interim R eport#13 Protocol Exygen P0001131; 3M Study Number E05-0210 Amendment 3 This amendment modifies the following portion(s) of the protocol: P :r o t o c o l r e a d s T Fe s t i n g a c i l i t y Exygen Research 3058 Research Drive State College, PA 15801 Phone: (814) 272-1039 A :m e n d t o r e a d T Fe s t in g a c il it ie s Exygen Research 3058 Research Drive State College, PA 15801 Phone: (814) 272-1039 3M Environmental Laboratory Building 2-3E-09 935 Bush Avenue St. Paul, MN 55106 Re a s o n : Addition of 3M Environmental Laboratory as a testing facility allows selected water samples to be sent to the 3M facility for analysis of PFBs , PFHS, and PFOS using the most current version of ETS 8-154 "Determination of Perfluorinated Acids, Alcohols, Amides, and Sulfonates in Water by Solid Phase Extractions and High Performance Liquid Chromatography/Mass Spectrometry". The following modifications to ETS 8-154.1 will be incorporated into the study: (1) The sample volume extracted and the final SPE elution volume may be adjusted, at the analyst's discretion, to better achieve the detection limits specified in study objectives. As written, ETS 8 154.1 calls for a 40 mL extraction volume with a 5 mL elution volume which results in an eight fold sample concentration. The actual volumes used and the resulting overall concentration factor will be given in the final report. Associated method quality control samples will demonstrate that the volume modifications do not impact method accuracy and precision. (2) A solvent (unextracted) calibration curve may be analyzed with the extracted calibration curve to ascertain extraction efficiency. Samples will be analyzed using the extracted curve unless otherwise stated in the final report. Data from the solvent calibration curve will not be included or discussed in the final report unless it is used to strengthen experimental observations/explanations. (3) ETS 8-154.1 makes no mention of sample surrogate spikes; however, all samples will be spiked with at least one of the following radiolabeled surrogates, PFOA [1,2 13C] (perfluorooctanoic acid), PFOS [18O2] and/or PFNA [1,2 13C] (perfluorononanoic acid - C9).The concentration of the surrogate spike may vary depending on the collection event, but typical samples are spiked with a nominal concentration of 0.05 ng/mL. Surrogate spike recoveries will be documented and discussed in the final report. Surrogate recoveries between 10025% will be deemed as meeting method criteria for accuracy. (4) Several method blanks may be prepared and analyzed to better determine a "representative" area count value for the method blank. The method blank area count is instrumental in determining the method limit of quantitation and understanding the average and range of area counts possible is critical. If a method blank value is used as a "zero point" in the final calibration curve, then a brief explanation in the final report or in the raw data as a Note to File must be E05-0210 GLP Protocol Amendment #3 Page 2 of 4 E05-0210 Interim R eport#13 Tennessee R iver Page 158 of 160 3M Environm ental Laboratory E05-0210 Interim R eport#13 Protocol Exygen P0001131; 3M Study Number E05-0210 Amendment 3 included to describe why that specific method blank was selected as the zero point (closest to the average value, most conservative (largest) value, etc.) (5) The overall analytical uncertainty for each target (non-surrogate) analyte will be estimated using both the method accuracy and precision. The method accuracy and precision will be calculated using data from laboratory contorl spike (LCS) samples prepared and analyzed with the sample set(s). Replicate LCSs must be prepared each day samples and/or other quality control samples are prepared. A sample calculation of how the analytical uncertainty was determined will be provided in the final report and/or raw data. A new revision of ETS 8-154 may incorporate all or several of the modifications listed above. If a new version of ETS 8-154 is issued during the course of this study, sample results must meet the new method requirements regardless if they are listed above. 3M Environmental Laboratory management will approve all documented deviations to the 3M Environmental Laboratory quality system (SOPs, methods, etc.) that occur during the course of this investigation. E05-0210 E05-0210 Interim R eport#13 GLP Protocol Amendment #3 Tennessee R iver Page 3 of 4 Page 159 of 160 3M Environm ental Laboratory E05-0210 Interim Report#13 Protocol Exygen P0001131; 3M Study Number E05-0210 Amendment 3 Amendment Approval MichaelA. Santoro, 3M , Sponsor Representative Date William K. Reagen, Ph.D. 3M Environmental Laboratory Manager Date Jaisimha Kesari, Weston Solutions, Study Director Date E05-0210 GLP Protocol Amendment #3 E05-0210 Interim Report#13 Tennessee River Page 4 of 4 Page 160 o f 160